HK1233672B - Integrated system for nucleic acid amplification and detection - Google Patents

Description

Integrated systems for nucleic acid amplification and detection

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent Application No. 61/953,316, filed on March 14, 2014.

Technical Field

The present teachings relate to designs and methods for obtaining biometric data and nucleic acids suitable for use in human identification and forensic science.

introduction

Forensic evidence and biometric data are often used together to identify perpetrators of criminal activity, as well as for identifying missing persons, victims of major disasters, paternity testing, and to exonerate innocent individuals. The ability to simultaneously collect an individual's biometric data (e.g., fingerprints, aperture or retinal scans, images, or photographs) and biological samples (e.g., forensic evidence, including but not limited to blood, tissue, hair, body fluids, or oral samples) can provide systems for accelerating the identification, access control, and screening of individuals. Furthermore, maintaining the identification of relevant data points and associating data with corresponding biological samples can be complex and prone to errors that can compromise the chain of custody. Therefore, a need remains for accurately collecting, correctly associating, and processing biometric data and biological samples from a single individual in one collection step or workflow.

Summary of the Invention

In a first aspect, a device for collecting a biological sample containing nucleic acid from an individual is provided, comprising a base substrate assembly having at least a first surface area, wherein the at least first surface area is transparent or translucent to energy waves capable of imaging at least one ridge and valley feature of an appendage of the individual; configured to collect the biological sample from the appendage; and made of one or more materials compatible with the reaction conditions of a nucleic acid amplification reaction. In some embodiments, the base substrate assembly can be a polymeric film or glass. In various embodiments, all surface areas of the substrate assembly can be transparent or translucent to the energy waves. In various embodiments, all surface areas of the base substrate assembly can be configured to be compatible with the nucleic acid amplification reaction conditions. In various embodiments, the at least first surface area can be hydrophilic. In some embodiments, the base substrate assembly further comprises a second surface area surrounding the at least first surface area, wherein the second surface area is hydrophobic.

The apparatus may further include a frame assembly for supporting the substrate assembly. In some embodiments, the frame assembly may be formed of a material selected from the group consisting of polymers, metals, metal alloys, glass, silicon materials, or combinations thereof.

The device may further include a mask assembly, wherein the mask assembly is configured to: be connected to the base substrate assembly; surround the at least first surface area of the base substrate assembly; and be made of one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction. In some embodiments, the mask assembly may allow for the addition of nucleic acid amplification reagents to the at least first surface area of the base substrate assembly. In various embodiments, after the biological sample from the appendage of the individual is collected, the mask assembly may be placed in position surrounding the at least first surface area of the base substrate assembly. In some embodiments, the mask assembly may be connected to the base substrate assembly by a hinge connection. In other embodiments, the mask assembly may be connected to the base substrate assembly by an adhesive. In various embodiments, the mask assembly may be made of a polymeric material.

The device may further include a cover component. In some embodiments, the cover component may include a material compatible with the reaction conditions of the nucleic acid amplification reaction.

In various embodiments, the base substrate assembly can be chemically modified with a chemical functional group selected from the group consisting of amino, chloromethyl, hydroxyl, carboxyl, and quaternary amino groups. In some embodiments, the chemical functional groups that modify the base substrate assembly can include a linking group selected from the group consisting of an amino-functionalized linking group, a sulfhydryl linking group, a homobifunctional linking group, and a heterobifunctional linking group. In some embodiments, the base substrate assembly can be chemically modified to covalently or non-covalently attach reagents, including but not limited to enzymes, binding partners, surfactants, anionic polymers, or zwitterionic substances. In some embodiments, the base substrate assembly can be configured to allow the release of the at least one nucleic acid from the biological sample. The base substrate assembly can be compatible with nucleic acid sequencing reaction conditions.

The device may further include a base substrate assembly having an identifier for associating the biological sample of the individual with the at least one ridge and valley characteristic of the individual, wherein the identifier is alphanumeric, graphical, magnetic, or electromagnetic. In other embodiments, when the mask assembly is present, the mask assembly may further include an identifier for associating the biological sample of the individual with the at least one ridge and valley characteristic of the individual, wherein the identifier is alphanumeric, graphical, magnetic, or electromagnetic. In some embodiments, the identifier may be a barcode.

The device may further be configured for archiving the biological sample or for shipping the sample to another location for testing.

In another aspect, a system for collecting a biological sample containing nucleic acid and at least one ridge and valley characteristic from an individual is provided, comprising: at least a first imaging assembly comprising a scanning surface configured to allow energy waves to penetrate the scanning surface, wherein the energy waves are configured to image the at least one ridge and valley characteristic of an appendage of the individual; and a device for collecting a biological sample containing nucleic acid from an individual, comprising a base substrate assembly having at least a first surface area, wherein the at least first surface area is transparent or translucent to energy waves capable of imaging the at least one ridge and valley characteristic of the appendage of the individual; configured to collect the biological sample from the appendage; and made of one or more materials compatible with the reaction conditions of a nucleic acid amplification reaction. In some embodiments, the base substrate assembly can be a polymeric film or glass. In some embodiments, the system is configured to collect the at least one ridge and valley characteristic by imaging the appendage through the scanning surface and the base substrate assembly of the device while the appendage is positioned on at least the first surface area of the base substrate assembly. In some embodiments, the base substrate assembly of the device of the system can be a polymeric film or glass. In various embodiments, all surface areas of the substrate assembly can be transparent or translucent to the energy waves. In various embodiments, all surface areas of the base substrate assembly can be configured to be compatible with the nucleic acid amplification reaction conditions. In various embodiments, the at least first surface area can be hydrophilic. In some embodiments, the base substrate assembly further includes a second surface area surrounding the at least first surface area, wherein the second surface area is hydrophobic.

The apparatus of the system may further include a frame assembly for supporting the substrate assembly. In some embodiments, the frame assembly may be formed of a material selected from the group consisting of polymers, metals, metal alloys, glass, silicon materials, or combinations thereof.

The apparatus of the system may further include a mask assembly, wherein the mask assembly is configured to: be coupled to the base substrate assembly; surround the at least first surface area of the base substrate assembly; and be made of one or more materials compatible with the reaction conditions of a nucleic acid amplification reaction. In some embodiments, the mask assembly may allow for the addition of nucleic acid amplification reagents to the at least first surface area of the base substrate assembly. In various embodiments, after the biological sample from the appendage of the individual is collected, the mask assembly may be placed in position surrounding the at least first surface area of the base substrate assembly. In some embodiments, the mask assembly may be coupled to the base substrate assembly by a hinge connection. In other embodiments, the mask assembly may be coupled to the base substrate assembly by an adhesive. In various embodiments, the mask assembly may be made of a polymeric material.

The device of the system may further include a cover component. In some embodiments, the cover component may include a material compatible with the reaction conditions of the nucleic acid amplification reaction.

In various embodiments, the base substrate assembly of the device of the system can be chemically modified with a chemical functional group selected from the group consisting of amino, chloromethyl, hydroxyl, carboxyl, and quaternary amino groups. In some embodiments, the chemical functional group modifying the base substrate assembly can include a linking group selected from the group consisting of an amino-functionalized linking group, a sulfhydryl linking group, a homobifunctional linking group, and a heterobifunctional linking group. In some embodiments, the base substrate assembly can be chemically modified to covalently or non-covalently attach reagents, including but not limited to enzymes, binding partners, surfactants, anionic polymers, or zwitterionic substances. In some embodiments, the base substrate assembly can be configured to allow the release of the at least one nucleic acid from the biological sample. The base substrate assembly can be compatible with nucleic acid sequencing reaction conditions.

The apparatus of the system may further comprise a base substrate assembly having an identifier for associating the biological sample of the individual with the at least one ridge and valley characteristic of the individual, wherein the identifier is alphanumeric, graphical, magnetic, or electromagnetic. In other embodiments, when the mask assembly is present, the mask assembly may further comprise an identifier for associating the biological sample of the individual with the at least one ridge and valley characteristic of the individual, wherein the identifier is alphanumeric, graphical, magnetic, or electromagnetic. In some embodiments, the identifier may be a barcode.

The device of the system may be further configured for archiving the biological sample or for shipping the sample to another location for testing.

In some embodiments, the scanning surface can be formed of a material selected from the group consisting of plastic, polyolefin, polystyrene, metal, metal alloy, glass, silicon, or combinations thereof. In some embodiments, the material of the scanning surface can be transparent or translucent. In various embodiments, the system can be configured to simultaneously collect the biological sample and the at least one ridge and valley feature.

The system may further include a processor configured to transmit the ridge and valley signatures of the individual to at least one database containing ridge and valley signatures, the database selected from the group consisting of: forensic, criminal, identity, child identity, missing persons, immigration, DMV, terrorist, paternity, arrestee, criminal database, access control, and any combination thereof.

The system may further include a nucleic acid amplification reaction assembly, wherein the reaction assembly may be configured to subject the device containing the biological sample on the base substrate assembly to a nucleic acid amplification reaction. In some embodiments, the reaction assembly may be configured to subject the device to thermal cycling conditions. In various embodiments, the nucleic acid amplification reaction on the device may be sealed with a sealing solution. In some embodiments, the nucleic acid amplification reaction may be part of an INDEL, SNP, or STR analysis. In some embodiments, the reaction assembly may be further configured to subject more than one device to a nucleic acid amplification reaction simultaneously. In various embodiments, the reaction assembly may be further configured to subject the device to a nucleic acid sequencing reaction.

The system can further include a detection component, wherein the detection component can be configured to detect at least one product of the nucleic acid amplification, thereby identifying the corresponding sequence of the at least one nucleic acid amplification product. In some embodiments, identification of the sequence of the at least one nucleic acid amplification product can provide nucleic acid sequence identification of the individual.

The system may further include a processor configured to transmit the nucleic acid sequence identity of the individual to at least one database containing nucleic acid sequence identities, the database selected from the group consisting of: forensic, criminal, identity, child identity, missing persons, immigration, Department of Motor Vehicles, terrorist, paternity testing, arrestee, criminal database, access control, and any combination thereof.

In yet another aspect, a kit is provided comprising a device for collecting a biological sample containing nucleic acid from an individual, the device comprising a base substrate assembly having at least a first surface area, wherein the at least first surface area is transparent or translucent to energy waves capable of imaging at least one ridge and valley feature of an appendage of the individual; configured to collect the biological sample from the appendage; and made of one or more materials compatible with the reaction conditions of a nucleic acid amplification reaction, and optionally instructions for use thereof.

In some embodiments, the base substrate assembly of the device of the kit can be a polymeric film or glass. In various embodiments, all surface areas of the substrate assembly can be transparent or translucent to the energy waves. In various embodiments, all surface areas of the base substrate assembly can be configured to be compatible with the nucleic acid amplification reaction conditions. In various embodiments, the at least first surface area can be hydrophilic. In some embodiments, the base substrate assembly further includes a second surface area surrounding the at least first surface area, wherein the second surface area is hydrophobic.

The device of the kit may further include a frame assembly for supporting the substrate assembly. In some embodiments, the frame assembly may be formed of a material selected from the group consisting of polymers, metals, metal alloys, glass, silicon materials, or combinations thereof.

The device of the kit may further include a mask assembly, wherein the mask assembly is configured to: be connected to the base substrate assembly; surround the at least first surface area of the base substrate assembly; and be made of one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction. In some embodiments, the mask assembly may allow for the addition of nucleic acid amplification reagents to the at least first surface area of the base substrate assembly. In various embodiments, after the biological sample from the appendage of the individual is collected, the mask assembly may be placed in position surrounding the at least first surface area of the base substrate assembly. In some embodiments, the mask assembly may be connected to the base substrate assembly by a hinge connection. In other embodiments, the mask assembly may be connected to the base substrate assembly by an adhesive. In various embodiments, the mask assembly may be made of a polymeric material.

The device of the kit may further include a cover component. In some embodiments, the cover component may include a material compatible with the reaction conditions of the nucleic acid amplification reaction.

In various embodiments, the base substrate assembly of the device of the kit can be chemically modified with a chemical functional group selected from the group consisting of amino, chloromethyl, hydroxyl, carboxyl, and quaternary amino groups. In some embodiments, the chemical functional groups that modify the base substrate assembly can include linking groups selected from the group consisting of amino-functionalized linking groups, sulfhydryl linking groups, homobifunctional linking groups, and heterobifunctional linking groups. In some embodiments, the base substrate assembly can be chemically modified to covalently or non-covalently attach reagents, including but not limited to enzymes, binding partners, surfactants, anionic polymers, or zwitterionic substances. In some embodiments, the base substrate assembly can be configured to allow the release of the at least one nucleic acid from the biological sample. The base substrate assembly can be compatible with nucleic acid sequencing reaction conditions.

The device of the kit may further comprise a base substrate assembly having an identifier for associating the biological sample of the individual with the at least one ridge and valley characteristic of the individual, wherein the identifier is alphanumeric, graphical, magnetic, or electromagnetic. In other embodiments, when the mask assembly is present, the mask assembly may further comprise an identifier for associating the biological sample of the individual with the at least one ridge and valley characteristic of the individual, wherein the identifier is alphanumeric, graphical, magnetic, or electromagnetic. In some embodiments, the identifier may be a barcode.

The device of the kit may further be configured for archiving the biological sample or for shipping the sample to another location for testing.

The kit may further include a reagent for stabilizing the biological sample on the substrate assembly for archiving or shipping. The kit may further include a reagent for amplifying the nucleic acid of the biological sample. The kit may further include a sealing solution. In some embodiments, the reagent may be suitable for performing at least one of: STR analysis, SNP analysis, and Indel analysis.

The kit may further include at least one package for protecting the device from contamination prior to use. The kit may further include at least one package for protecting the device from contamination while archiving or shipping after the nucleic acid is deposited on the base substrate assembly. In some embodiments, the at least one package may include an identifier for associating the device containing the biological sample with at least one of the ridge and valley signatures of the individual and the identity of the individual providing the biological sample.

In another aspect, a method for collecting a biological sample containing at least one nucleic acid and collecting at least one ridge and valley characteristic of an appendage of an individual is provided, comprising the steps of providing at least a first imaging assembly. The at least first imaging assembly is configured to provide an energy wave capable of imaging the at least one ridge and valley characteristic; and has a scanning surface configured to allow the energy wave to penetrate the scanning surface. The method further comprises the steps of providing an apparatus having a base substrate assembly having at least a first surface area, wherein the base substrate assembly is configured to allow the energy wave to penetrate the substrate; collecting the biological sample from the appendage of the individual; and being made of one or more materials configured to be compatible with the reaction conditions of a nucleic acid amplification reaction. The apparatus can be any of the embodiments of the apparatus described herein having any combination of components. The method further comprises the steps of placing the appendage of the individual on the scanning surface and in contact with the base substrate assembly, thereby collecting the biological sample; and collecting the at least one ridge and valley characteristic from the appendage imaged by the energy wave. The appendage imaged can be a finger, a toe, a palm, or a sole of a foot. The steps of collecting the biological sample and collecting the at least one ridge and valley characteristic can be performed simultaneously.

In some embodiments, the at least first imaging component may be an optical scanner or a capacitive scanner, and the at least one ridge and valley feature may be collected electronically.

The method may further include the steps of transmitting the at least one ridge and valley signature of the individual to at least one database containing ridge and valley signatures, the database selected from the group consisting of: forensic, criminal, identity, child identity, missing persons, immigration, motor vehicle department, terrorist, paternity testing, arrestee, criminal database, access control and any combination thereof.

The method may further include the step of providing an identifier on at least one component of the device to associate the biological sample of the individual with the at least one ridge and valley characteristic of the individual, wherein the identifier is alphanumeric, graphical, magnetic, or electromagnetic. In some embodiments, the identifier is a barcode.

The method may further comprise the step of archiving at least the base substrate assembly of the device after performing the step of collecting the biological sample.

The method may further comprise the step of mailing the device to another location for archiving or testing after performing the step of collecting the biological sample.

The method may further require the step of subjecting the biological sample collected on the base substrate assembly to a nucleic acid amplification reaction. In some embodiments, the nucleic acid amplification reaction is a polymerase chain reaction. The method may further require the step of performing a nucleic acid sequencing reaction while the biological sample is still present on the base substrate assembly. In some embodiments, the nucleic acid amplification reaction and the nucleic acid sequencing reaction can be performed in the same reaction well without transferring or stopping between processes. In some embodiments, the amplification reaction can be a component of an STR analysis, a SNP analysis, or an Indel analysis.

The method may further require the step of placing a mask assembly of the device on the base substrate assembly, thereby surrounding at least the first surface area of the base substrate assembly. The mask assembly may be made of one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction. In some embodiments, the step of placing the mask assembly on the base substrate assembly is performed before the step of collecting the biological sample. In other embodiments, the step of placing the mask assembly on the base substrate assembly is performed after the step of collecting the biological sample and before the step of subjecting the biological sample on the base substrate assembly to the nucleic acid amplification reaction. The method may further include the step of sealing the nucleic acid amplification reaction with a sealing solution. The reaction well may not have any other upper closure besides the sealing solution. Alternatively, the method may include the step of placing a cover assembly of the device on the biological sample collected on the base substrate assembly during the nucleic acid amplification reaction. In some embodiments, the step of placing the cover assembly on the biological sample on the base substrate assembly is performed after the reagents for the nucleic acid amplification reaction have been added to the biological sample on the base substrate assembly and before the reaction begins. The method may further include the step of placing a cover assembly on the biological sample collected on the base substrate assembly for shipping or archiving prior to nucleic acid amplification. The method may further include the step of detecting the sequence of the nucleic acid, thereby identifying the individual. The method may further include the step of transmitting the nucleic acid sequence identity of the individual to at least one database containing nucleic acid sequence identities, the database selected from the group consisting of: forensic, criminal, identity, child identity, missing persons, immigration, DMV, terrorist, paternity, arrestee, criminal database, access control, and any combination thereof.

In yet another aspect, a method for identifying an individual is provided, comprising providing at least a first imaging component configured to provide an energy wave capable of imaging at least one ridge and valley feature of an appendage of the individual and comprising a scanning surface configured to allow the energy wave to penetrate the scanning surface. The method further comprises providing an apparatus comprising a base substrate assembly having at least a first surface area, wherein the substrate assembly is configured to allow the energy wave to penetrate the substrate; collecting a biological sample comprising nucleic acid from the appendage of the individual; and being made of one or more materials configured to be compatible with reaction conditions of a nucleic acid amplification reaction. The apparatus may be any of the embodiments of the apparatus described herein having any combination of components. The method further comprises the steps of positioning the appendage of the individual on the scanning surface and in contact with the at least first surface area of the base substrate assembly, thereby collecting the biological sample; collecting the at least one ridge and valley feature from the appendage imaged by the energy wave, thereby providing identification of the ridge and valley feature of the individual; subjecting the nucleic acid of the biological sample collected on the base substrate assembly to a nucleic acid amplification reaction; and detecting the sequence of the nucleic acid, thereby identifying the individual. The steps of collecting the biological sample and collecting the at least one ridge and valley feature may be performed simultaneously.

In some embodiments, the at least first imaging component can be an optical scanner or a capacitive scanner. In various embodiments, the at least one ridge and valley feature can be collected electronically. In some embodiments, the appendage can be a finger, toe, palm, or sole. In other embodiments, the nucleic acid amplification reaction can be a polymerase chain reaction. In various embodiments, the method can include the step of performing a DNA sequencing reaction while still present on the base substrate assembly. In some embodiments, the amplification reaction can be a component of an STR analysis, a SNP analysis, or an Indel analysis.

The method may further include placing a mask assembly of the device on the base substrate assembly so as to surround at least the first surface area of the substrate, wherein the mask assembly is made of one or more materials compatible with the reaction conditions of a nucleic acid amplification reaction. In some embodiments, placing the mask assembly on the base substrate assembly may be performed before collecting the biological sample. In other embodiments, placing the mask assembly on the base substrate assembly may be performed after collecting the biological sample and before subjecting the biological sample on the substrate assembly to a nucleic acid amplification reaction.

The method may further comprise the step of sealing the nucleic acid amplification reaction mixture with a sealing solution.The nucleic acid amplification reaction mixture may not have other upper packaging.

The method may further comprise the step of placing a cover assembly of the device over the biological sample collected on the base substrate assembly. The cover assembly may be placed over the at least first surface of the base substrate assembly containing the nucleic acid after the biological sample is collected to facilitate shipping and archiving the device. In some embodiments, the cover assembly may be placed over the biological sample on the substrate assembly after reagents for the nucleic acid amplification reaction have been added to the biological sample on the base substrate assembly and before the reaction begins.

The method may further include the steps of transmitting the at least one ridge and valley signature of the individual to at least one database containing ridge and valley signatures, the database selected from the group consisting of: forensic, criminal, identity, child identity, missing persons, immigration, motor vehicle department, terrorist, paternity testing, arrestee, criminal database, access control and any combination thereof.

The method may further include the step of providing an identifier on at least one component of the device to associate the biological sample of the individual with the at least one ridge and valley characteristic of the individual, wherein the identifier is alphanumeric, graphical, magnetic, or electromagnetic. In some embodiments, the identifier is a barcode.

The method may further comprise the step of archiving at least the base substrate assembly of the device after performing the step of collecting the biological sample.

The method may further comprise the step of mailing the device to another location for archiving or testing after performing the step of collecting the biological sample.

The method may further comprise the step of transmitting the nucleic acid sequence identity of the individual to at least one database comprising nucleic acid sequence identities, the database being selected from the group consisting of forensic, criminal, identity, child identity, missing persons, immigration, DMV, terrorist, paternity, arrestee, criminal database, access control, and any combination thereof.

The method may further comprise the step of collecting at least a second image of the individual, the at least second image being selected from a facial image, an iris image, a retinal image, or an ear image of the individual. In some embodiments, the identifier may associate the at least second image with the biological sample and the at least one ridge and valley characteristic marker of the individual.

In another aspect, a system for identifying a target nucleic acid is provided, comprising: a base substrate assembly comprising one or more hydrophilic sites separated by hydrophobic regions, wherein at least one hydrophilic site is configured to contain at least one target nucleic acid, and further wherein the base substrate assembly is made of one or more materials compatible with the reaction conditions of a nucleic acid amplification reaction; a reaction assembly configured to amplify the at least one target nucleic acid at the at least one hydrophilic site to form at least one nucleic acid amplification product; and a detection assembly configured to identify at least one nucleic acid sequence of the at least one nucleic acid amplification product. In some embodiments, the base substrate assembly can be a polymeric film or glass. In various embodiments, the reaction assembly may not require a solid upper seal assembly for the substrate. The reaction assembly can be configured to provide an oil seal to the at least one hydrophilic site for conducting the nucleic acid amplification reaction. The reaction assembly can be configured to provide an oil seal to each of the one or more hydrophilic sites. The reaction assembly can be configured to provide nucleic acid amplification reagents to the at least one target nucleic acid contained at the at least one hydrophilic site on the base substrate assembly. The reaction component can be configured to provide suitable reaction conditions for nucleic acid amplification of the at least one target nucleic acid contained on the at least one hydrophilic site on the base substrate component. The system can be further configured to extract a volume of the at least one nucleic acid amplification reaction product from under the oil seal for use in identifying the at least one nucleic acid sequence using the detection component.

The detection component of the system can be selected from a fluorescent dye sequencing component, a semiconductor sequencing component, or a pyrophosphate sequencing component. In some embodiments, the detection component can be a fluorescent dye sequencing component. In some embodiments, the fluorescent dye sequencing component can be a mobility-dependent fluorescent dye sequencing component. In some embodiments, the mobility-dependent fluorescent dye sequencing component can be capillary electrophoresis.

In yet another aspect, the present invention provides a base substrate assembly for nucleic acid amplification and identification, comprising: one or more hydrophilic sites separated by hydrophobic regions, wherein at least one hydrophilic site is configured to contain at least one target nucleic acid; wherein the base substrate assembly is made of one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction; and further wherein the base substrate assembly is configured to allow extraction of an aliquot of the amplified target nucleic acid for detection of the nucleic acid sequence identity. The base substrate assembly can be a polymer or glass.

In yet another aspect, a kit is provided, comprising a base substrate assembly for nucleic acid amplification and identification, the base substrate assembly comprising: one or more hydrophilic sites separated by hydrophobic regions, wherein at least one hydrophilic site is configured to contain at least one target nucleic acid; wherein the base substrate assembly is made of one or more materials compatible with the reaction conditions of a nucleic acid amplification reaction; and further wherein the base substrate assembly is configured to allow extraction of an aliquot of the amplified target nucleic acid for detection of the nucleic acid sequence identity, and optionally instructions for use thereof. The kit may further comprise reagents for a nucleic acid amplification reaction. The kit may further comprise reagents for a nucleic acid sequencing reaction.

In another aspect, a method for identifying at least one target nucleic acid is provided, comprising the following steps: providing a base substrate assembly having one or more hydrophilic sites separated by hydrophobic regions, wherein at least one hydrophilic site comprises at least one target nucleic acid, and further wherein the base substrate assembly is made of one or more materials compatible with the reaction conditions of a nucleic acid amplification reaction; providing a reagent suitable for amplifying the at least one target nucleic acid at the at least one hydrophilic site; performing a nucleic acid amplification reaction on the at least one target nucleic acid to form at least one nucleic acid amplification product; and detecting the nucleic acid sequence of the at least one nucleic acid amplification product, thereby identifying the at least one target nucleic acid.

In some embodiments, a solid upper sealing component may not be provided for the substrate. In various embodiments, the method further comprises the steps of providing a sealing solution layer to the at least one hydrophilic site containing the at least one nucleic acid for the nucleic acid amplification reaction. In some embodiments, the sealing solution layer may be provided to each of the one or more hydrophilic sites. In some embodiments, the method further comprises the steps of providing a nucleic acid amplification reagent to the at least one target nucleic acid contained on the at least one hydrophilic site on the substrate. The method may further comprise providing suitable reaction conditions for performing nucleic acid amplification on the at least one target nucleic acid contained on the at least one hydrophilic site on the base substrate assembly. In some embodiments, a certain volume of the at least one nucleic acid amplification reaction product is extracted from under the oil seal to identify the at least one nucleic acid sequence by the detection component.

In some embodiments of the method, the detection component can be selected from a fluorescence detection component, a semiconductor detection component, or a pyrophosphate detection component. In other embodiments, the detection can be a fluorescent dye sequencing component. In other embodiments, the fluorescent dye sequencing component can be a mobility-dependent fluorescent dye sequencing component. In other embodiments, the mobility-dependent fluorescent dye sequencing component can be capillary electrophoresis.

On the other hand, the present invention also provides a system for identifying target nucleic acids, which includes (a) a reaction cartridge, which includes: a dissolution well configured to accommodate a sample substrate having a biological sample, the sample substrate including the target nucleic acid and a dissolution buffer of an aliquot, wherein the biological sample is dissolved to release the target nucleic acid; a PCR well configured to accommodate a PCR reagent of an aliquot, wherein the released target nucleic acid is amplified to form an amplified target nucleic acid sample; a sealing solution well configured to accommodate a sealing solution of an aliquot, wherein the sealing solution is transferred to the PCR well to seal the open PCR; and optionally, a well configured to transfer the aliquot. The transfer end of any one of the released target nucleic acid, the sealing solution and the amplified target nucleic acid sample; and optionally, a detection sample preparation hole operably connected to the reaction barrel, wherein the detection sample preparation hole is configured to accommodate an aliquot of the sample configured to prepare a reagent for identifying the nucleic acid sequence of the amplified target nucleic acid sample; and wherein the reaction barrel contains one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction; (b) a reaction component that is configured to amplify the released target nucleic acid to form the amplified target nucleic acid sample; and (c) a detection component that is configured to identify the nucleic acid sequence of the amplified target nucleic acid sample. In some embodiments, the reaction barrel can be made of a polymer or glass. In some embodiments, the sample substrate can be a section of paper substrate, including but not limited to paper threading from a paper substrate on which a biological sample has been collected, or a cheek swab. In various embodiments, the reaction barrel may not require a solid upper closure component for the reaction barrel. In some embodiments, the system can be configured to transfer an aliquot of the released target nucleic acid from the dissolution hole to the PCR hole. In certain embodiments, the reaction assembly can be configured to provide suitable reaction conditions to carry out nucleic acid amplification to the target nucleic acid of the release contained in the PCR wells of the reaction barrel. In various embodiments, the system can be configured to extract a certain volume of the amplified target nucleic acid sample from under the oil seal and transfer it to the detection sample preparation hole to prepare the amplified target nucleic acid sample for identifying the nucleic acid sequence by the detection assembly. In other embodiments, the detection assembly can be selected from a fluorescent dye sequencing assembly, a semiconductor sequencing assembly or a pyrophosphate sequencing assembly. In certain embodiments, the detection assembly can be a fluorescent dye sequencing assembly. In certain embodiments, the fluorescent dye sequencing assembly can be a mobility-dependent fluorescent dye sequencing assembly. In certain embodiments, the mobility-dependent fluorescent dye sequencing assembly can be a capillary electrophoresis assembly. The system can further include a plurality of reaction barrels.

On the other hand, a reaction cartridge for nucleic acid amplification and identification is provided, comprising (a) a dissolution well configured to accommodate a sample substrate having a biological sample, the sample substrate comprising the target nucleic acid and a dissolution buffer of an aliquot, wherein the biological sample dissolves to release the target nucleic acid; (b) a PCR well configured to accommodate a PCR reagent of an aliquot, wherein the released target nucleic acid is amplified to form an amplified target nucleic acid sample; (c) a sealing solution well configured to accommodate a sealing solution of an aliquot, wherein the sealing solution is transferred to the PCR well to seal the open PCR; and optionally, (d) a transfer end configured to transfer any one of the released target nucleic acid, the sealing solution, and the amplified target nucleic acid sample of the aliquot; and optionally, (e) a detection sample preparation well operably connected to the reaction cartridge, wherein the detection sample preparation well is configured to accommodate an aliquot of a reagent configured to prepare the sample for identifying the nucleic acid sequence of the amplified target nucleic acid sample; and further wherein the reaction cartridge is made of one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction. In some embodiments, the reaction cartridge is made of one or more polymers, glass, or a combination thereof.

On the other hand, a kit is provided, which includes a reaction cartridge for nucleic acid amplification and identification, the reaction cartridge including (a) a dissolution well configured to accommodate a sample substrate having a biological sample, the sample substrate including the target nucleic acid and an aliquot of a dissolution buffer, wherein the biological sample is dissolved to release the target nucleic acid; (b) a PCR well configured to accommodate an aliquot of PCR reagents, wherein the released target nucleic acid is amplified to form an amplified target nucleic acid sample; (c) a sealing solution well configured to accommodate an aliquot of a sealing solution, wherein the sealing solution is transferred to the PCR well to seal the open PCR; and optionally, (d) a transfer end configured to transfer any one of the released target nucleic acid, the sealing solution and the amplified target nucleic acid sample of the aliquot; and optionally, (e) a detection sample preparation well operably connected to the reaction cartridge, wherein the detection sample preparation well is configured to accommodate an aliquot of a reagent configured to prepare the sample for identifying the nucleic acid sequence of the amplified target nucleic acid sample; and further wherein the reaction cartridge is made of one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction, and optionally instructions for use thereof. In some embodiments, the reaction cartridges of the batch are made of one or more polymers, glass, or a combination thereof. In various embodiments, the test kit may further include reagents for nucleic acid amplification reactions. In various embodiments, the test kit may further include a sealing solution. In some other embodiments, the test kit may further include reagents for nucleic acid sequencing reactions.

On the other hand, a method for identifying a target nucleic acid is provided, comprising the following steps: (a) providing a reaction cartridge, the reaction cartridge comprising: a dissolution well configured to accommodate a sample substrate containing a biological sample, the sample substrate comprising the target nucleic acid and an aliquot of a dissolution buffer, wherein the biological sample is dissolved to release the target nucleic acid; a PCR well configured to accommodate an aliquot of a PCR reagent, wherein the released target nucleic acid is amplified to form an amplified target nucleic acid sample; a sealing solution well configured to accommodate an aliquot of a sealing solution, wherein the sealing solution is transferred to the PCR well to seal the open PCR; and optionally, a transfer well configured to transfer any of the released target nucleic acid, the sealing solution, and the amplified target nucleic acid sample. end; and optionally, a detection sample preparation hole operably connected to the reaction barrel, wherein the detection sample preparation hole is configured to accommodate an aliquot of a reagent configured to prepare the sample for identifying the nucleic acid sequence of the amplified target nucleic acid sample; and wherein the reaction barrel includes one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction; (b) providing a reagent suitable for amplifying the released target nucleic acid in the PCR hole of the reaction barrel; (c) sealing the PCR hole with an aliquot of the sealing solution; (d) performing a nucleic acid amplification reaction on the released target nucleic acid to form the amplified target nucleic acid sample; and (e) detecting the nucleic acid sequence of the amplified target nucleic acid sample, thereby identifying the at least one target nucleic acid. In some embodiments, the reaction barrel can be a polymeric film or glass. In some embodiments, the sample substrate can be a section of paper substrate or a cheek swab. In various embodiments, the method can further include the steps of adding the sample substrate including the biological sample containing the target nucleic acid to the dissolution hole, and dissolving the biological sample to release the target nucleic acid. In other embodiments, the method may further include the step of transferring an aliquot of the released target nucleic acid from the dissolution well to the PCR well using the transfer tip after the biological sample has been dissolved to release the target nucleic acid. In various embodiments, a solid upper sealing assembly may not be provided for the substrate. In some embodiments, the method may further include the step of transferring the sealing solution using the transfer tip to the PCR well of the reaction cartridge containing the released nucleic acid and the aliquot of PCR reagent. In some embodiments, the method may further include the step of providing suitable reaction conditions to amplify the released target nucleic acid contained in the PCR well of the reaction cartridge to form the amplified target nucleic acid sample. In some embodiments, a volume of the amplified target nucleic acid sample may be extracted from beneath the oil seal using the transfer tip and transferred to the test sample preparation well. In other embodiments, the detection assembly may be selected from a fluorescent dye sequencing assembly, a semiconductor sequencing assembly, or a pyrophosphate sequencing assembly. In other embodiments, the detection assembly may be a fluorescent dye sequencing assembly. In some embodiments, the fluorescent dye sequencing assembly may be a mobility-dependent fluorescent dye sequencing assembly. In some embodiments, the mobility-dependent fluorescent dye sequencing assembly may be a capillary electrophoresis assembly. In some embodiments, the reagents configured to prepare the aliquot of the sample for identifying the nucleic acid sequence of the amplified target nucleic acid sample may include a denaturant. The denaturant may be formamide. In some embodiments, the reagents of the aliquot further include a size standard.

In the following description, certain aspects and embodiments will become apparent. It should be understood that a given embodiment does not necessarily have all aspects and features described herein. It should be understood that these aspects and embodiments are merely illustrative and explanatory and do not limit the present invention.

There remains a need for improved systems, kits, and methods for collecting fingerprint, toe print, palm print, or sole print, and biological sample data for the purpose of identifying and verifying the identity of a human individual.

BRIEF DESCRIPTION OF THE DRAWINGS

1A and 1B illustrate a system for simultaneously collecting appendicular ridge and valley features and a biological sample from an individual, according to various embodiments.

FIG2 illustrates a device for simultaneously collecting adjacent ridge and valley signatures and a biological sample from an individual, and also demonstrates its compatibility for nucleic acid amplification without transferring the biological sample.

3 is a graphical representation of STR analysis performed on nucleic acid collected from an individual using the device of FIG. 2 .

4 illustrates another embodiment of a device for collecting ridge and valley signatures and biological samples from an individual that is compatible with nucleic acid amplification.

FIG. 5 is a graphical representation of a fingerprint image obtained using the apparatus of FIG. 4 .

6 is a graphical representation of STR analysis performed on nucleic acid collected from an individual using the device of FIG. 4 .

7 illustrates another embodiment of a device for collecting ridge and valley signatures and biological samples from an individual that is compatible with nucleic acid amplification.

8A and 8B illustrate two embodiments of devices for collecting ridge and valley signatures and biological samples from an individual that are compatible with nucleic acid amplification.

9 is a graphical representation of STR analysis performed on nucleic acid collected from an individual using the device of FIG. 8A .

10 illustrates another embodiment of a device for collecting ridge and valley signatures and biological samples from an individual that is compatible with nucleic acid amplification.

FIG. 11 is a graphical representation of a fingerprint image obtained using the apparatus of FIG. 10 .

12 is a graphical representation of STR analysis performed on nucleic acid collected from an individual using the device of FIG. 10 .

13 illustrates another embodiment of a device for collecting ridge and valley signatures and biological samples from an individual that is compatible with nucleic acid amplification.

14A and 14B illustrate another embodiment of a device for collecting ridge and valley signatures and a biological sample from an individual that is compatible with nucleic acid amplification, and also show assembly of the device after collection of the ridge and valley signatures and the biological sample.

15 is a graphical representation of STR analysis of nucleic acid collected from an individual using a variation of the device of FIGS. 14A and 14B .

16 is a graphical representation of STR analysis performed on nucleic acid collected from an individual using another variation of the device of FIGS. 14A and 14B .

17A and 17B illustrate another embodiment of a device for collecting ridge and valley signatures and a biological sample from an individual that is compatible with nucleic acid amplification, and also show assembly of the device after collection of the ridge and valley signatures and the biological sample.

FIG18 illustrates a base substrate assembly of an integrated amplification and detection system having selected hydrophilic reaction sites filled with open PCR samples.

19 is a graphical representation of STR analysis of amplified nucleic acids using the base substrate assembly of FIG. 18 .

20 is a graphical representation of STR analysis of amplified nucleic acids using the base substrate assembly of FIG. 18 .

Figure 21 is a diagrammatic representation of the reaction components of an integrated amplification and identification system.

DETAILED DESCRIPTION

For the purpose of interpreting this specification, the following definitions will apply and, whenever appropriate, terms used in the singular will also include the plural and vice versa. Unless the contrary meaning is clearly intended (e.g., in the document in which the term is initially used), for the purpose of interpreting this specification and its associated claims, in the event that any definition set forth below conflicts with the usage of the term in any other document, including any document incorporated herein by reference, the definition set forth below shall always prevail. It should be noted that, as used in this specification and the appended claims, the singular forms "a", "an" and "the" include plural referents unless expressly and unambiguously limited to one referent. Unless otherwise stated, the use of "or" means "and/or". The use of "comprise", "comprises", "comprising", "include", "includes" and "including" is interchangeable and is not intended to be limiting. Furthermore, when the term “comprising” is used in the description of one or more embodiments, those skilled in the art will appreciate that, in some specific cases, the embodiments may alternatively be described using the language “consisting essentially of” and/or “consisting of.”

As used herein, "DNA" and "nucleic acid" are used interchangeably.

As used herein, "oligonucleotide" and "polynucleotide" are interchangeable and generally refer to polymers of nucleotide subunits of about 200 base pairs or less in size.

As used herein, a "biological sample" refers to a component derived from within or on the body of an individual.

As used herein, "body fluid" refers to fluid derived from within the body of an individual.

As used herein, "digital imaging device" refers to a device that is capable of digitizing an image of an object.

As used herein, "DNA sequencing" refers to determining the sequence identity of nucleotides in a DNA molecule.

As used herein, "filter paper" refers to semi-permeable paper.

As used herein, "housing" refers to a structure that at least partially surrounds a device and enables physical movement or performance of physical actions including, but not limited to, lighting, scanning, and the like.

As used herein, an "identifier" refers to a tag that can be used to catalog/associate similarly tagged data or data from a single source.

As used herein, "image capture device" refers to a type of camera or scanning device.

As used herein, "imaging assembly" refers to a device capable of at least one of: capturing, developing, storing, retrieving, and transmitting an image.

As used herein, "Indel" refers to an insertion or deletion of a nucleic acid, usually DNA, within a nucleic acid sequence.

As used herein, "isolated" refers to separation of nucleic acids from either or both naturally occurring materials or environmental chemicals/materials.

As used herein, "light emitting diode" refers to an LED, a semiconductor light source.

As used herein, "multispectral illuminator" refers to multiple frequencies/wavelengths across the electromagnetic spectrum used to capture image data.

As used herein, "open PCR" refers to a nucleic acid polymerase chain reaction amplification performed in a reaction vessel that does not have a solid upper package. The reaction is performed under a sealing layer that isolates the reaction mixture from the environment. The sealing layer can be suitably added to the reaction mixture of the target nucleic acid and the PCR reagents, or the sealing layer can be added to the target nucleic acid and the PCR reagents are added to the target nucleic acid through the sealing layer. Typically, the sealing layer is a liquid, including but not limited to oil, such as mineral oil.

As used herein, "optically collecting" refers to obtaining data (eg, ridge and valley features) or images by illuminating the data or image and capturing the result.

As used herein, "appendage" refers to one of the distal-most parts of a limb and includes a finger or toe.

As used herein, "camera" refers to LED cameras, digital cameras, still cameras, video cameras, and virtual cameras.

As used herein, "polymerase chain reaction" or PCR is the amplification of nucleic acids and consists of an initial denaturation step to separate the strands of a double-stranded nucleic acid sample, followed by repeated steps of: (i) an annealing step that allows the amplification primers to specifically anneal to positions flanking the target sequence; (ii) an extension step that extends the primers in the 5' to 3' direction, thereby forming an amplicon polynucleotide complementary to the target sequence; and (iii) a denaturation step that causes the amplicon to separate from the target sequence (Mullis et al., eds., The Polymerase Chain Reaction, Birk Hauser, Boston, Mass. (1994)). Each of the above steps can be performed at different temperatures, preferably using an automated thermal cycler (Applied Biosystems LLC, a division of Life Technologies Corporation, Foster City, CA.). If desired, RNA samples can be converted to DNA/RNA heteroduplexes or duplex cDNA by methods known to those skilled in the art. The PCR method also includes reverse transcriptase-PCR and other reactions that follow the PCR principle.

As used herein, "ridge and valley signatures" refer to friction ridges, also known as surface contours, on the palmar surfaces of the fingers, the surfaces of the palms and hands, and the toe surfaces of the feet and toes. Specifically, the friction ridges provided by indentation made by imaging one or more fingers are referred to as fingerprints.

As used herein, "SNP analysis" refers to the assessment of the presence or absence of single nucleotide polymorphism (SNP) markers following amplification of the loci containing the SNP markers.

As used herein, "STR analysis" refers to the assessment of the alleles of short tandem repeat (STR) markers following amplification of the loci containing the STR markers.

As used herein, "sequentially" refers to the order in which steps are performed.

As used herein, "topological indentation" refers to the ridge and valley surface shape of a finger, hand, toe, or foot.

Reference will now be made to various embodiments, examples of which are illustrated in the accompanying drawings.

There is a need for improvements in collecting biological samples containing nucleic acids while also obtaining biometric information (e.g., fingerprints). Specifically, it would be advantageous to obtain biological samples non-invasively. It has been found that, under suitable conditions, when the ridge and valley signatures of an individual's appendages are imaged through a substrate and a scanning surface, sufficient nucleic acid is deposited when the appendages contact the substrate to provide a DNA profile with a signal sufficient to identify the individual. In some embodiments, devices, systems, and methods are disclosed for analyzing biological samples that, after being deposited on a substrate, are further processed, such as to allow direct amplification of nucleic acids without transferring the nucleic acid or biological sample to any other container or receiving surface. Processing of the nucleic acids collected on the substrate itself (including nucleic acid amplification, nucleic acid sequencing reactions, and optionally dissolution) allows direct PCR amplification in extremely small volumes, which can provide increased sensitivity and a simplified workflow. Further simplification has been achieved by performing nucleic acid amplification directly on the substrate (i.e., performing open PCR) without the need for a solid cover for the reaction wells, and by using a sealing solution layer on the mixture of amplification reagents and deposited nucleic acids. Additional manipulation of the collected nucleic acid may not be required, further preventing the loss of the limited amount of nucleic acid provided in the biological sample.

The devices, systems, kits, and methods described herein provide the ability to directly amplify fingerprint nucleic acid from the same substrate on which the fingerprint was collected. This improvement enhances chain-of-custody integrity and reduces the potential for sample handling errors that may occur when transferring fingerprint nucleic acid from the substrate on which the fingerprint nucleic acid was deposited to a PCR reaction vessel.

The improvements provided by the open PCR format provide amenability to automation and allow for easy recovery of PCR products by simple pipetting through the sealing solution layer.

Furthermore, the small thermal cycler footprint and weight make it an ideal PCR module for fully integrated STR typing or other identification systems. The AmpliSpeed slide thermal cycler used in this study weighs only 2.9 kg and measures only approximately 126 mm x approximately 295 mm x approximately 112 mm.

System. A system for collecting a biological sample containing nucleic acid and at least one ridge and valley characteristic from an individual is described herein. The system comprises at least a first imaging assembly having a scanning surface configured to allow energy waves to penetrate the scanning surface, wherein the energy waves are configured to image the at least one ridge and valley characteristic of the individual's appendage; and a device having a base substrate assembly having at least a first surface area, wherein the at least first surface area is transparent or translucent to the energy waves capable of imaging the at least one ridge and valley characteristic of the individual's appendage; configured to collect the biological sample from the appendage; and made of one or more materials compatible with the reaction conditions of a nucleic acid amplification reaction. The system is configured to collect the at least one ridge and valley characteristic by imaging the appendage through the scanning surface and the base substrate assembly of the device while the appendage is positioned on at least the first surface area of the base substrate assembly. The biological sample comprising at least one nucleic acid is collected from the skin of the appendage of the individual. Skin is a complex organ and comprises more than one layer of cells and tissue types. The epidermis refers to the tissue on the surface of human or animal skin and includes substances secreted therefrom or derived from the underlying layers of the skin, from which DNA useful for identifying an individual can be readily obtained. The outermost layer of the skin is the stratum corneum. Below the epidermis of the skin is the dermis, which contains fibroblasts, macrophages, and adipocytes, each of which has a nucleus containing nucleic acids. In addition, the dermis has a vascular network of blood veins, arteries, and lymphatic vessels containing white blood cells with nuclei, as do the arrector pili muscle tissue, sebaceous glands, and body fluids present in the dermis. In addition, the sweat glands and blood vessels in the dermis can contain genetic material in the form of intact cells or free DNA, which can be released to the epidermis for collection by the systems and methods of the present invention. Any or all of these substances containing nucleic acids are encompassed by biological samples. In various embodiments of the present invention, collecting biological samples is non-invasive.

The system can collect at least one ridge and valley signature and a biological sample containing nucleic acid of an individual simultaneously with at least a first imaging component, or can collect the signature and the biological sample sequentially, or vice versa, collect the biological sample and the signature sequentially. The collection method enables the collection of both the biological sample and the at least one ridge and valley signature while the individual can touch only one device. No additional action or step of touching an additional platen or substrate may be required to collect the biological sample and the at least one ridge and valley signature. A ridge refers to a friction ridge, a raised portion of the epidermis layer of the skin of a finger, toe, palm, or sole of the foot, and a valley is a depression in the epidermis between two adjacent ridges. Ridges and valleys are often referred to as fingerprints, palm prints, toe prints, or footprints, depending on the source of the ridge and valley signature.

The system may further include a nucleic acid amplification reaction assembly, wherein the reaction assembly is configured to allow the device containing the biological sample on the base substrate assembly to undergo a nucleic acid amplification reaction. In some embodiments in which the system provides a reaction assembly, the reaction assembly provides a sealing solution for the nucleic acid amplification reaction. Using the sealing solution to isolate the reaction mixture from the environment without the need for a physical cover for the reaction wells can provide a streamlined workflow. In some embodiments, the system includes a nucleic acid reaction assembly further configured to allow the device to undergo a nucleic acid sequencing reaction.

The system can further include a component wherein the detection component is configured to detect at least one nucleic acid amplification product, thereby identifying the corresponding sequence of the at least one nucleic acid amplification product. In some embodiments, identification of the sequence of the at least one nucleic acid amplification product provides nucleic acid sequence identification of the individual.

The system may further include a processor configured to transmit the ridge and valley signatures of the individual to at least one database containing ridge and valley signatures, the database selected from the group consisting of: forensic, criminal, identity, child identity, missing persons, immigration, DMV, terrorist, paternity, arrestee, criminal database, access control, and any combination thereof. The system may further include a processor configured to transmit the nucleic acid sequence identity of the individual to at least one database containing nucleic acid sequence identities, the database selected from the group consisting of: forensic, criminal, identity, child identity, missing persons, immigration, DMV, terrorist, paternity, arrestee, criminal database, access control, and any combination thereof.

In various embodiments, the system uses at least a first imaging system to simultaneously collect at least one ridge and valley feature of an individual and a biological sample containing nucleic acid, or to collect the feature and the biological sample sequentially, or vice versa, to collect the biological sample and the feature sequentially, while requiring the individual to touch the device only once.

At least a first imaging component. In various embodiments, the system includes at least a first imaging component configured to obtain ridge and valley features of an individual. The imaging component can utilize energy waves, such as light or other energy waves as described above (e.g., electromagnetic, capacitive, infrared, or acoustic (e.g., ultrasonic)-based components) to provide images of the ridge and valley features. When the term imaging component is used in the context of capturing ridge and valley features, this includes any component that captures a digital or analog electronic representation of the ridge and valley features.

Ridge and valley feature signatures can also be obtained using a non-contact three-dimensional ridge and valley scanner utilizing digital processing components. (Wang, Yongchang; Q. Hao, A. Fatehpuria, D. L. Lau and L. G. Hassebrook (2009). “Data Acquisition and Quality Analysis of 3-Dimensional Fingerprints”. Florida: IEEE conference on Biometrics, Identity and Security. http://vis.uky.edu/~realtime3d/Doc/3D_Fingerprint_Quality.pdf. Retrieved March 2010. Wang, Yongchang; D. L. Lau and L. G. Hassebrook (2010). “Fit-sphere unwrapping and performance analysis of 3D Fingerprints”. Applied Optics. pp. 592-600).

In one embodiment of the present teachings, the system can include at least a first optical imaging assembly. In other embodiments, the system can include at least a first solid-state ridge and valley feature reader. The optical imaging assembly has an illumination component for optically collecting the ridge and valley feature using an optical scanner as is known to those skilled in the art. The optical scanner can be an array of multiple light emitting diodes or a multispectral illuminator. The optical scanner can be a camera, an active pixel imager, a CMOS imager, an imager that images in multiple wavelengths, a CCD camera, a photodetector array, or a TFT imager. In the optical scanner, a beam of light is passed through the scanning surface to illuminate the topological impression obtained by the appendage (including a finger, hand, palm, toe, sole, or foot) when placed against the scanning surface of at least the first imaging assembly.

Any suitable instrument can be used to obtain the image of the attachment according to the method described herein. Some instruments and techniques include but are not limited to US4537484, US6175407, US6665427, US8014581, US8036431, US5177353, US6282303, US 6188781, US6741729, US6122394, US6826000, US6496630, US6628813, US6983062, US7162060, US7164440, US7657067, US8073209, US7190817, US755841 0. US7565541, US7995808, US7899217, US7890158, US7835554, US7831072, US7819311, US7804984, US7801339, US7801338, US7751594, US7735729, US76 68350, US7627151, US7620212, US7613504, US7545963, US7539330, US7508965, US7460696, US7440597, US7394919, US7386152, US7347365, US7263213, US7203345, US7147153, US6816605, US6628809, US6560352, US20110235872, US20110211055, US20110165911, US20110163163, US20110085708, US2010 0246902, US20100067748, US20090245591, US20090148005, US20090092290 , US20090080709, US20090046903, US20080304712, US20080298649, US2008 0297788, US20080232653, US20080192988, US20080025580, US20080025579 , US20070116331, US20070030475, US20060274921, US20060244947, US2006 0210120, US20060202028, US20060110015, US20060062438, US20060002598, US20060002597, US20050271258, US20050265586, US20050265585, US20050205667, US20050185847, US20050007582, US20040240712, US20040047493, US20030223621, US20030078504, US20020183624, or US20020009213.

Scanning Surface. At least the first imaging assembly includes a scanning surface configured to allow energy waves to penetrate the scanning surface to image at least one ridge and valley feature of an accessory of an individual disposed on at least a portion of the scanning surface. The scanning surface can be transparent or translucent to the energy waves used to image the at least one ridge and valley feature. The scanning surface can be formed from a material selected from the group consisting of plastic, polyolefin, polystyrene, metal, metal alloy, glass, silicon, or combinations thereof. In some embodiments, the surface can comprise a derivatized plastic, a derivatized polyolefin, a derivatized polystyrene, a derivatized metal, a derivatized metal alloy, a derivatized glass, a derivatized silicon, or a combination of materials.

Device. A device for collecting a biological sample containing nucleic acid from an individual is provided, wherein the device comprises a base substrate assembly having at least a first surface region, wherein the at least first surface region is transparent or translucent to energy waves capable of imaging at least one ridge and valley feature of an appendage of the individual; is configured to collect the biological sample from the appendage; and is made of one or more materials compatible with reaction conditions commonly used in nucleic acid amplification reactions. The materials may further be compatible with nucleic acid sequencing reaction conditions.

A base substrate assembly. The base substrate assembly comprises at least a first surface area, wherein the at least first surface area is transparent or translucent to energy waves capable of imaging at least one ridge and valley feature of an appendage of an individual; is configured to collect a biological sample containing nucleic acid from the appendage; and is made of one or more materials compatible with the reaction conditions of a nucleic acid amplification reaction. After the biological sample containing nucleic acid is collected on the base substrate assembly, the nucleic acid deposited on the substrate can be subjected to a nucleic acid amplification reaction, thus eliminating the need for transfer or additional handling of the nucleic acid. In situ amplification simplifies workflow, reduces the likelihood of sample mishandling, and increases the probability of obtaining consistent amplification results from limited sample quantities, while also obtaining the biological sample in a single, non-invasive procedure. The base substrate assembly can be any suitable polymeric film or glass. In some embodiments, at least the first surface area of the base substrate assembly can be transparent or translucent to energy waves capable of imaging at least one ridge and valley feature. In other embodiments, all surface areas of the base substrate assembly can be transparent or translucent to energy waves. In some embodiments, the base substrate assembly is substantially transparent to energy waves. In other embodiments, all surface areas of the base substrate assembly can be compatible with the reaction conditions of a nucleic acid amplification reaction. In various embodiments, at least the first surface area of the base substrate assembly can be hydrophilic. In other embodiments, at least the first surface area of the base substrate assembly can be treated with a PCR-compatible adhesive to facilitate capture of biological samples. In various embodiments, the base substrate assembly can further include a second surface area surrounding the first surface area. The second surface area of the base substrate assembly can be hydrophobic.

In FIG1A , a finger 101 is placed on a base substrate assembly 110A disposed on a scanning surface to collect a biological sample containing nucleic acids and ridge and valley features onto the substrate 110A. In some embodiments, the fingerprint containing the biological sample containing nucleic acids is deposited within a first surface region 110B-1, which may be approximately 20 mm wide ( FIG1B ). The surrounding surface region 110B-2 may have the same surface features as the first surface region 110B-1, or it may be modified to have different properties.

Polymeric Film Base Substrate Assembly. In various embodiments, the base substrate assembly can be a polymeric film. In some embodiments, the polymeric film can be dimensionally stable enough to maintain structural integrity on the scanned surface. In other embodiments, the polymeric film can be attached to or connected to a carrier. Suitable polymeric films include natural polymeric materials, including but not limited to starch, agarose, alginates, carrageenan, or synthetic polymer gels, or mixtures thereof.

Synthetic polymers can also be used as the base substrate component. To form a polymeric film that can be used as the base substrate component, all types of polymerization can be used to synthesize the polymer gel that forms part or all of the base substrate component, including cationic, anionic, copolymeric, chain copolymeric, cross-linked, non-cross-linked polymerization, etc. Essentially any type of polymer or copolymer that can be formed from a fluid precursor can be incorporated as part or all of the base substrate component, including but not limited to homopolymers, random copolymers, triblock polymers, radial polymers, linear polymers, branched polymers, and graft copolymers. An exemplary, non-limiting list of suitable polymers includes cellulosic polymers, including but not limited to hydroxyethylcellulose (HEC), hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC); sugar derivatives, including but not limited to dextran, mannitol, and glucopyranosides; pluronic copolymer liquid crystals; polyurethanes; polyacids, including but not limited to lactic acid and acrylic acid; polyamides; polyacrylamides, including but not limited to unsubstituted, N-substituted, and N,N-disubstituted acrylamides; polycarbonates; polyacetylene; polydiacetylene; polyphosphazenes; polysiloxanes; polyolefins; polyesters; polyethers; poly(etherketones); poly(alkylene oxides), including but not limited to polyethylene oxide (PEO), polyethylene glycol (PEG), and polypropylene oxide (PPO); poly(ethylene terephthalate); poly(methyl methacrylate); polystyrene; substituted polystyrenes, including but not limited to polystyrene sulfonate (PSS) and polyanethole (PASA); poly(vinyl pyrrolidone); proteinaceous materials, and/or combinations and/or copolymers thereof.

In some embodiments, the polymeric film base substrate assembly can be selected from commercially available films, including but not limited to 3M PP2200, 3M PP2500, 3M CG600, or 3M CG3300. Many types of commercially available films can be suitable for use as substrates without interfering or substantially interfering with the fingerprint recognition process. Suitable films have the ability to retain DNA on the film when an individual's appendage contacts the film to image the ridge and valley features. Other suitable membranes allow for direct amplification of DNA from a collected biological sample in the presence of the membrane. Still other suitable membranes allow for at least one nucleic acid from a biological sample to be rinsed from the surface of the membrane and subsequently amplified.

In some embodiments, the polymeric film base substrate assembly does not modify the polymeric film.

Glass Base Substrate Assembly. In other embodiments, the base substrate assembly can be made of glass, and can be made of any suitable glass material that allows energy waves to penetrate the substrate to image the ridge and valley features of an individual's appendages, collect a biological sample, and is compatible with nucleic acid amplification reaction conditions. Many types of commercially available glass can be suitable for use as the base substrate assembly without interfering or substantially interfering with the fingerprint recognition process. Suitable glasses have the ability to capture DNA on the glass when the individual's appendages contact the glass to image the ridge and valley features. Some suitable glasses allow the collected biological sample containing DNA to be transferred to another substrate. In some embodiments, the glass substrate allows the transfer of at least one nucleic acid of the biological sample from the base substrate assembly while retaining other components of the biological sample. Other suitable glasses allow the DNA of the collected biological sample to be amplified directly in the presence of the glass. Still other suitable glasses allow the collected biological sample containing DNA to be rinsed from the surface of the glass and subsequently amplified.

Surface modification of polymeric or glass base substrate assemblies. The base substrate assembly can have one or more modified surface regions. At least a first surface region of the base substrate assembly can be treated with a PCR-compatible adhesive to aid in the collection of biological samples, or it can be hydrophilic. The glass base substrate assembly can be treated with a reagent to create a hydrophobic surface region surrounding the surface region that is contacted by the individual's finger during fingerprint imaging and deposits the biological sample. The hydrophobic second surface can be obtained by treating the glass with a reagent including, but not limited to, a silanizing agent. The hydrophobic second surface region of the base substrate assembly can help retain nucleic acid amplification reagents within the first surface region containing the deposited nucleic acid during the amplification reaction. In some embodiments, the base substrate assembly is modified to be adhesive. The adhesive can be PCR-compatible. In other embodiments, the polymeric film is viscosity-enhanced to aid in the collection of biological samples without causing physical handling difficulties.

In some embodiments, the base substrate component can be modified with chemically reactive groups to allow ionic, covalent, or hydrogen bonding interactions with substances present in the biological sample. Suitable chemically reactive groups that can modify the base substrate component include, but are not limited to, thiols, amines, carboxylic acids, and the like, as generally known in the art. Some examples of chemical modifications include, but are not limited to, amino groups, including aliphatic and aromatic amines; carboxylic acids; aldehydes; amides; chloromethyl groups; hydrazides; hydroxyls; sulfonates; and phosphates. These functional groups can be used to add any number of modifications to the polymeric or glass base substrate component, typically using known chemistries (including, but not limited to, amino-functionalized linking groups, sulfhydryl linking groups, and the like). There are a variety of sulfhydryl-reactive linking groups known in the art, such as SPDP, maleimide, α-haloacetyl, and pyridyl disulfide. Similarly, amino groups on the polymeric or glass base substrate component can be linked using linking groups; for example, many stable bifunctional groups are well known in the art, including homobifunctional and heterobifunctional linking groups. In another embodiment, the carboxyl groups on the base substrate component can be derivatized using well-known linking groups. For example, carbodiimides activate carboxyl groups for attack by good nucleophiles such as amines.

In some embodiments, chemical modification of the base substrate assembly is performed to covalently or non-covalently attach reagents to aid in the collection of DNA from a subject, including but not limited to enzymes, binding partners, surfactants, anionic polymers, or zwitterionic substances. In some embodiments, the chemical modification can introduce a conductive polymer to the base substrate assembly to aid in the collection of DNA from a subject. If the conductive polymer forms part of the base substrate assembly, the system can further provide a suitable current across the base substrate assembly to provide iontophoresis assistance when the accessory is in contact with the base substrate assembly and the circuit is closed, thereby driving charged nucleic acids from the accessory to the base substrate assembly.

In other embodiments, when the base substrate assembly is chemically modified to covalently attach one or more reagents, an enzyme (including but not limited to proteinase K) can be attached to the base substrate assembly to facilitate nucleic acid extraction from the appendage or from a collected biological sample. In other embodiments, the enzyme is covalently attached to the base substrate assembly and can be a protease selected from the group consisting of keratinase, papain, bromelain, and proteinase K. In other embodiments, the base substrate assembly is covalently modified to attach a charged polymer to facilitate nucleic acid extraction from the appendage or from a collected biological sample. In other embodiments, a surfactant is non-covalently attached to the base substrate assembly to facilitate nucleic acid extraction from the appendage or from a collected biological sample. In some embodiments, a zwitterionic substance can be present in the base substrate assembly to facilitate separation of nucleic acids from other components of the collected cells. In some other embodiments, the base substrate assembly is chemically modified to covalently attach an antibody to a target moiety present on the skin of the appendage or in the collected biological sample, thereby providing a binding partner for the target moiety. In some embodiments in which the binding partner is immobilized on the base substrate assembly, the binding partner can undergo cleavage either together with or separately from its bound target moiety.

Frame Assembly. The apparatus may further include a frame for the base substrate assembly. In some embodiments, the apparatus may be configured to allow collection of at least one ridge and valley feature through the frame and base substrate assembly. The frame may be made of any material capable of forming a solid frame. Suitable materials for the frame may have any of a variety of properties, depending on the specific embodiment, including, for example, porosity, non-porousness, rigidity, elasticity, flexibility, toughness, and/or chemical resistance to one or more solvents commonly used in the reactions described herein. In some embodiments, the frame is formed from a material selected from the group consisting of a polymer, a metal, a metal alloy, glass, a silicon material, or combinations thereof. Suitable polymers include, but are not limited to, plastics; polypropylene, polyethylene, polybutylene, polyurethane, nylon, polymers such as acrylic, acrylonitrile butadiene styrene (ABS), ULTEM (polyetherimide), acetal copolymers, PROPYLUX HS (heat stabilized polypropylene), RADEL A (polyethersulfone), RADEL R (polyarylethersulfone), UDEL (polysulfone), NORYL PPO (polyphenylene oxide and styrene), polycarbonate, UHMW-PE (ultra-high molecular weight polyethylene), polyetheretherketone (PEEK), polyphenylene sulfide (PPS, TECHTRON or RYTON), polyolefins, or polystyrene; metals such as aluminum, iron, steel, or alloys; other materials such as glass or silicon, or derivatives or combinations of these or other suitable materials. In some embodiments of the present teachings, the frame is made of a transparent or translucent material.

Mask assembly. The device may include a mask assembly configured to be connected to a base substrate assembly. The connection may be permanent, including but not limited to bonding with an adhesive or a hinge connection that allows repositioning after the biological sample is deposited. The connection of the mask assembly to the base substrate assembly may be performed after the biological sample is deposited onto the base substrate assembly. The mask assembly may surround at least a first surface area of the base substrate assembly, while making at least the first surface area of the base substrate assembly available for adding nucleic acid amplification reaction reagents, a sealing layer, and sampling for detection, among other functions. The mask assembly may have through holes to create PCR reaction holes after bonding to the base substrate assembly, wherein the through holes surround at least the first surface area of the base substrate assembly where the nucleic acid has been deposited. The mask assembly may be made of one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction. The mask assembly may be made of one or more polymeric materials or glass materials.

Polymer Mask Assembly. In various embodiments, the base substrate assembly can be a polymer. Suitable polymers include natural polymeric materials including, but not limited to, starch, agarose, alginate, carrageenan, or synthetic polymer gels or mixtures thereof.

Synthetic polymers can also be used as mask components. To form polymers that can be used as mask components, all types of polymerization can be used to synthesize the polymer gel that forms part or all of the mask component, including cationic, anionic, copolymeric, chain copolymeric, cross-linked, non-cross-linked polymerization, etc. Essentially any type of polymer or copolymer that can be formed from a fluid precursor can be incorporated as part or all of the mask component, including but not limited to homopolymers, random copolymers, triblock polymers, radial polymers, linear polymers, branched polymers, and graft copolymers. An exemplary, non-limiting list of suitable polymers includes cellulosic polymers, including but not limited to hydroxyethylcellulose (HEC), hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC); sugar derivatives, including but not limited to dextran, mannitol, and glucopyranosides; pluronic copolymer liquid crystals; polyurethanes; polyacids, including but not limited to lactic acid and acrylic acid; polyamides; polyacrylamides, including but not limited to unsubstituted, N-substituted, and N,N-disubstituted acrylamides; polycarbonates; polyacetylene; polydiacetylene; polyphosphazenes; polysiloxanes; polyolefins; polyesters; polyethers; poly(etherketones); poly(alkylene oxides), including but not limited to polyethylene oxide (PEO), polyethylene glycol (PEG), and polypropylene oxide (PPO); poly(ethylene terephthalate); poly(methyl methacrylate); polystyrene; substituted polystyrenes, including but not limited to polystyrene sulfonate (PSS) and polyanethole (PASA); poly(vinyl pyrrolidone); proteinaceous materials, and/or combinations and/or copolymers thereof. The mask component can be natural or synthetic rubber, latex, or vinyl polymer.

In some embodiments, the polymer mask assembly can be selected from commercially available films, including but not limited to 3M PP2200, 3MPP2500, 3M CG600, or 3M CG3300. Many types of commercially available polymers can be suitable for use as the mask assembly without interfering or substantially interfering with the fingerprint recognition process. Some suitable polymers have the ability to retain DNA on the polymer when an individual's appendage contacts the polymer while imaging ridge and valley features. Other suitable polymers allow DNA from a collected biological sample to be amplified directly in the presence of the polymer. Still other suitable polymers allow at least one nucleic acid from a biological sample to be rinsed from the surface of the polymer mask assembly and subsequently amplified.

In some embodiments, the polymeric mask assembly does not modify the polymeric film.

Glass Mask Assembly. In other embodiments, the mask assembly can be made of glass, and can be made of any suitable glass material that will not interfere with imaging the ridge and valley features of the individual's appendages and is compatible with the nucleic acid amplification reaction conditions. Many types of commercially available glass can be suitable for use as the mask assembly without interfering or substantially interfering with the fingerprint recognition process. Some suitable glasses have the ability to capture DNA on the glass during imaging of the ridge and valley features. Other suitable glasses allow DNA from a collected biological sample to be amplified directly in the presence of the glass. Still other suitable glasses allow a collected biological sample containing DNA to be rinsed from the surface of the glass and subsequently amplified.

Surface modification of a polymer or glass mask assembly. The mask assembly can have one or more surface regions having a modification. The lower surface of the mask can have an adhesive for bonding to the base substrate assembly, wherein bonding can occur before or after imaging of the ridge and valley features and deposition of the biological sample. The upper surface of the mask can have adhesive properties to collect additional amounts of biological sample, as portions of the finger contacting the base substrate assembly outside of at least the first surface region can deposit additional nucleic acid. The upper surface of the mask can have an adhesive for bonding to another layer to create a combined mask/cover assembly. Alternatively, the mask assembly can have an adhesive for bonding to a ring assembly that helps hold nucleic acid amplification reagents in place during open PCR. The upper surface of the mask assembly can be treated to be hydrophobic to hold the nucleic acid amplification reaction in place during in situ open PCR of nucleic acids on the base substrate assembly. When the mask assembly is glass, the upper surface of the glass mask assembly can be treated with a reagent to create a hydrophobic surface region surrounding the surface region where nucleic acid has been deposited. The hydrophobic surface can be obtained by treating the glass with a reagent including, but not limited to, a silanizing agent. The hydrophobic second surface region of the base substrate assembly can help retain nucleic acid amplification reagents within the first surface region containing the deposited nucleic acids during open PCR.

Cover assembly. The apparatus may further include a cover assembly configured to protect the base substrate assembly, the mask assembly, or both before and after collecting the ridge and valley features and the biological sample. In some embodiments, the cover assembly may also be present during the nucleic acid amplification reaction. In other embodiments, for example, when performing open PCR, the cover assembly is absent and the reaction mixture is protected from the environment by a sealing solution layer.

The cover assembly can be made of any suitable material (polymer or glass). In some embodiments, the cover assembly is transparent or translucent. In other embodiments, the cover assembly is made of a material compatible with PCR conditions. In some embodiments, the cover assembly has an adhesive on its lower surface so as to be bonded to the mask assembly to form a combination mask/cover assembly, which can define reaction holes for performing in situ PCR on the base substrate assembly. When the combination mask/cover assembly defines the reaction holes, it is aligned with at least the first surface area of the base substrate assembly, allowing reagents to be added to the nucleic acids deposited in at least the first surface area of the base substrate assembly. In some embodiments, access ports can be formed in the combination mask/cover assembly for delivering reagents and extracting sample aliquots for detection.

Any of the base substrate assembly, mask assembly, substrate frame, or cover assembly can have an identifier that associates a biological sample of an individual with at least one ridge and valley feature obtained from the individual.

Exemplary Device Examples. Exemplary devices are described that allow for direct amplification of fingerprint nucleic acids on a surface where nucleic acid has been collected using an open PCR format. Such devices integrate the functions of fingerprint collection from a finger, nucleic acid sample collection, and PCR amplification of the fingerprint nucleic acid using an open PCR format on the same device, without requiring any form of nucleic acid sample transfer. The geometry, number, and device dimensions of the fingerprint nucleic acid deposition areas described herein are for illustrative purposes only and are not intended to limit the scope of the present invention.

FIG2 illustrates a device 201 having a polymeric film base substrate assembly. The polymeric film substrate can be made from a variety of polymeric films, and in this particular embodiment, the substrate is a transparent adhesive film, wherein the adhesive is compatible with PCR conditions and reagents and can be configured to fit within the scanning window of an optical fingerprint scanner. The area of the base substrate assembly used for each fingerprint imaging/nucleic acid collection is approximately 2.5 cm by approximately 5 cm. One or more fingerprints can be imaged while each finger is in contact with the substrate. As shown in FIG2 , a reaction chamber having a diameter of approximately 8 mm can be formed on each fingerprint by placing a SecureSeal ^™ 8-hole mask (203) (Grace Bio-labs, AB8R-0.5) on top of the adhesive film. Each surface area containing nucleic acids (210-1.1, 210-1.2, 210-1.3, 210-1.4, 210-1.5, 210-1.6, 210-1.7, and 210-1.8) remains open and available for reagent addition. Each cavity formed by surrounding each fingerprint with a mask can be filled with a volume of approximately 50 μl of PCR reaction mixture. The cavity can be sealed on top of the mask using a second portion of a transparent adhesive film (not shown in FIG2 ). The assembled fingerprint PCR device can be placed on a flat thermal cycler block (e.g., a 9700 thermal cycler (Applied Biosystems)) and can be secured to the thermal block for thermal cycling.

Another embodiment of an apparatus 401 for collecting fingerprint images, fingerprint nucleic acid, and performing in situ nucleic acid amplification is shown in FIG4 . In this embodiment, the apparatus comprises three component layers and is shown in a closed configuration. The bottom layer is a transparent substrate 410 made of a material that inhibits thermal cycling during PCR. The top surface of the bottom layer may be hydrophobic, except for a hydrophilic circle indicated by area 410-1 (which may be approximately 10 mm in diameter). An intermediate layer mask 403 has 10 mm diameter through-holes that align with the hydrophilic circle 410-1 on the base substrate layer 401. Mask 403 has two functions. First, it allows fingerprint nucleic acid collection within the circular area 410-1 on the base substrate layer 410, while preventing fingerprint DNA collection on the hydrophobic areas of the top surface of the base substrate layer 401. Second, the intermediate layer allows fingerprint nucleic acid to be collected from the portion of the finger that contacts mask 403 surrounding the circular area 410-1 during fingerprint collection.

Top layer 404 acts as a protective covering for the middle and bottom layers and is removable to facilitate fingerprint acquisition and reagent addition. Cover 404 can be placed in place while handling the collected fingerprint nucleic acid or during transportation or mailing of the collected fingerprint nucleic acid to a different location for testing or archiving. Cover 404 can be present or removed during the nucleic acid amplification reaction performed on the nucleic acid collected by contact between the finger and the substrate during fingerprint imaging. When cover 404 is absent during nucleic acid amplification, the nucleic acid amplification reaction mixture generated by adding reagents to the nucleic acid present in hydrophilic region 410-1 can be sealed with a sealing solution layer, which can be an oil.

To use the device 401, it is placed on the scanning surface of a fingerprint imaging assembly, the cover 404 is removed and the hydrophilic circular area 410-1 is used to simultaneously collect a fingerprint image and nucleic acid from a finger placed in contact with the substrate for fingerprint imaging.

Many variations of this embodiment can be used in any combination. In another embodiment, the entire upper surface of base substrate 410 can be hydrophobic. The upper surface of base substrate 410 can be concave, forming shallow dimples, or any other desired shape to help retain the PCR reaction mixture and/or sealing solution during PCR. A hydrophilic circle can be located at the bottom of the dimple.

Mask layer 403 can be transparent. Mask layer 403 can also be translucent, as long as it does not interfere with fingerprint scanning. If the degree of transparency differs between the area within circular region 410-1 and the rest of the imaged area during the fingerprint scan, the fingerprint scanning software can make adjustments to obtain a fingerprint image of appropriate quality. Fingerprint nucleic acids collected on mask layer 403 can also be retained and archived for subsequent analysis, if necessary. In some embodiments, the upper surface of mask layer 403 can be covered with an adhesive to facilitate additional nucleic acid collection.

Another embodiment of a device is shown in FIG7 . Device 701 can contain two component layers. Base substrate layer 710 can be transparent or translucent and made of a material that can withstand the thermal cycles of PCR. Region 710-2 of the upper surface of base substrate layer 710 is hydrophobic and surrounds hydrophilic region 710-1, which is shaped like an oval in this embodiment. FIG7 is drawn to show two distinct regions of the upper surface of the base substrate as viewed through translucent upper cover layer 704. Hydrophilic region 710-1 can have dimensions of approximately 16 mm by approximately 20 mm. Hydrophilic region 710-1 is used to collect fingerprint nucleic acid from a finger used to image an individual's fingerprint.

Top layer 704 acts as a protective covering for the middle and bottom layers and is removable to facilitate fingerprint acquisition and amplification of nucleic acids collected in hydrophilic region 710-1. Cover 704 can be placed in place while handling the collected fingerprint nucleic acid or during transportation or mailing of the collected fingerprint nucleic acid to a different location for testing or archiving. Cover 704 can be removed during the nucleic acid amplification reaction of nucleic acids collected by contact between the finger and the substrate during fingerprint imaging. The nucleic acid amplification reaction mixture at 710-1 can be sealed with a sealing solution layer, which can be an oil.

To use the device 701, it is placed on the scanning surface of a fingerprint imaging assembly, the cover 704 is removed and the hydrophilic region 710-1 is used to simultaneously collect a fingerprint image and nucleic acid from a finger placed in contact with the substrate for fingerprint imaging.

Many variations of this embodiment can be used in any combination. In another embodiment, the entire upper surface of base substrate 710 can be hydrophobic. The upper surface of base substrate 710 can be concave, forming a shallow dimple, or any other desired shape to help retain the PCR reaction mixture and/or the sealing solution during PCR. The hydrophilic region 710-1 can be located at the bottom of the dimple.

Another embodiment of the device is shown in Figure 8A. This embodiment 801A illustrates the feasibility of capturing DNA from a fingertip on a PCR-compatible substrate and then forming a PCR reaction chamber directly on the fingerprint on the substrate using a combined mask and cover assembly 805A to perform direct nucleic acid amplification and optionally sequencing reactions. In one embodiment, nucleic acid from a finger contacting a base substrate layer 810A is collected in area 810A-1 on a transparent adhesive film (Life Technologies, 4306311). A reaction chamber on the fingerprint deposit 802A having a diameter of about 8 mm and a depth of about 1 mm is formed by placing a SecureSeal well (Grace Bio-labs, AB8R-0.5) on the area of the fingerprint nucleic acid deposit 802A in the enclosed area 810A-1 of the adhesive film 810A-2. The resulting chamber can be filled with 50 μL of PCR reaction mixture, and the top of the chamber can be sealed with another blank piece of transparent adhesive film. The assembled fingerprint PCR device can be thermally cycled on an AB9700 flat thermal cycler block to amplify nucleic acids and can include sequencing reactions to provide amplicons suitable for detection, which can include fluorescent or other types of labels for sequencing reactions that require labeling; or the amplification products can be detected by detection methods that do not require labeled amplicons (e.g., semiconductor sequencing).

Many variations of this embodiment are possible. The base substrate layer 810A may be free of any adhesive, and the lower surface of the combined mask/cover assembly 805A forming the reaction wells may instead have an adhesive compatible with PCR conditions and reagents to connect to the base substrate layer 810A to form an airtight reaction vessel surrounding the fingerprint nucleic acid deposit 802A in region 810A-1.

Another embodiment of a device is shown in FIG8B . Device 801B comprises a base substrate assembly 810B, wherein fingerprint nucleic acid 802B is deposited in region 810B-1, which is surrounded by region 810B-2 where no nucleic acid is deposited. A combined cover and mask 805B forms a reaction well in region 810B-1 and has two ports 806-1 and 806-2 extending through cover/mask 805B for adding reagents and extracting amplified nucleic acid for detection. An adhesive is applied to the base substrate layer surrounding fingerprint nucleic acid deposit 802B on the lower surface outside the well-forming region of combined cover/mask 805B or in region 810B-2.

Another embodiment of a device configured to capture nucleic acid from a finger while simultaneously performing fingerprint imaging is shown in FIG10 . Device 1001 is compatible with PCR reaction conditions and allows for nucleic acid amplification reactions, which may further include in situ nucleic acid identification (including but not limited to STR analysis) without the need to transfer the nucleic acid to another reaction well or reaction surface. The PCR reaction may be "open PCR," in which the PCR is covered only by a sealing solution to prevent evaporation during PCR thermal cycling.

Device 1001 comprises two component layers. Base substrate layer assembly 1010 is transparent and made of a material that can withstand PCR thermal cycling. The upper surface of base substrate layer assembly 1010 is hydrophobic, except for a hydrophilic region 1010-1, which may be approximately 10 mm in diameter, for collecting fingerprint DNA. The 10 mm diameter hydrophilic circular region shown with respect to device 1001 is merely illustrative and is not intended to exclude other shapes and sizes. Upper mask assembly layer 1003-1 has a through hole, approximately 10 mm in diameter, that aligns with hydrophilic region 1010-1 on base substrate layer assembly 1010. This region forms a reaction well for in situ PCR amplification of nucleic acids deposited within region 1010-1. Open PCR can be performed on nucleic acids deposited within region 1010-1, and the reaction can be sealed with a sealing solution during thermal cycling.

Many variations of this embodiment are possible. Region 1010-1 can be coated with a PCR-compatible adhesive to facilitate fingerprint DNA collection. Alternatively, the entire top surface of base substrate layer assembly 1010 can be hydrophobic or hydrophilic. Base substrate layer assembly 1010 can have a concave depression at region 1010-1 or any other desired shape to help retain the PCR reaction mixture and/or sealing solution during PCR. The fingerprint collection area can be located at the bottom of the depression.

Mask assembly layer 1003-1 can allow for the collection of fingerprint DNA from the portion of the finger outside the 10 mm aperture during the fingerprint collection process. The upper surface of mask assembly layer 1003-1 can be coated with an adhesive. The top layer can be transparent. Mask assembly layer 1003-1 can also be translucent; if so, the translucent mask assembly layer 1003-1 should not interfere with fingerprint scanning. If the degree of transparency varies between areas inside and outside area 1010-1 during fingerprint scanning, the fingerprint scanning software can be adjusted to obtain a high-quality fingerprint image. The fingerprint DNA collected on mask assembly layer 1003-1 can be archived and subsequently analyzed as needed. If necessary, a cover layer can be added to act as a protective layer.

Another embodiment of the device is shown in Figure 13. Device 1301 is made of a material that can inhibit the thermal cycling of PCR. The upper surface of the base substrate region 1301 of the device is hydrophobic (region 1310-2), except for the hydrophilic region 1310-1, which can be configured as an oval with dimensions of approximately 16 mm x approximately 20 mm for collecting fingerprint DNA. The dimensions described are only an example and are not intended to exclude other shapes and sizes. The device is placed on the scanning surface of the imaging assembly, and the individual places a finger in contact with the hydrophilic region 1310-1 to simultaneously deposit a biological sample containing nucleic acid and image the fingerprint through the base substrate layer assembly 1310 at region 1310-1. The collected nucleic acids can be amplified in situ using open PCR, wherein the sealing solution prevents evaporation of PCR reagents and/or solvents.

In other variations, the 16×20 mm hydrophilic oval area 1310-1 can instead be coated with a PCR-compatible adhesive to facilitate fingerprint DNA collection. Alternatively, the entire upper surface of device 1301 can be hydrophobic or hydrophilic. Device 1301 can have concave wells or other desired shapes to help retain the PCR reaction mixture and/or seal the solution during PCR. The fingerprint collection area can be located at the base of the well.

Another embodiment of the device is shown in FIG14 . Device 1401A is shown in assembly and has a base substrate assembly 1410A made of a film compatible with both fingerprint collection and PCR reaction conditions. The film of the upper surface area (including areas 1410A-1 and 1410A-2) can be coated with a PCR-compatible adhesive. The fingerprint image and fingerprint nucleic acid 1402A are collected into area 1410A-1 while the device is positioned on the scanning surface of the imaging assembly. Mask layer assembly 1403A with through-hole 1407A is another part of the device. Open PCR device 1401B is shown in its completed form in FIG14B , formed by bonding mask assembly layer 1403B, which can be a thin plastic or glass slide (e.g., 1 mm), with a 10 mm diameter through-hole 1407B, to base substrate layer assembly 1410B containing fingerprint nucleic acid 1402B still aligned with the opening of through-hole 1407B. Open PCR can then be performed on the fingerprint nucleic acid in situ and sealed with a sealing solvent layer (including but not limited to an oil layer). Detection of the amplified nucleic acid in the aliquot can be performed by extracting the aliquot from under the sealing solvent layer.

In one variation of the device illustrated in Figures 14A-B, base substrate assembly 1410A is a sheet of transparent adhesive film (Life Technologies, 4306311). The open PCR device 1401B is formed by bonding a glass mask assembly layer 1403B having a 10 mm diameter through hole to the adhesive film of base substrate assembly layer 1410A containing the fingerprint nucleic acid deposited at region 1410A-1. In another variation, the upper surface of glass mask assembly layer 1403B is hydrophobic to help retain the sealing solution during the open PCR reaction. In another variation, base substrate assembly 1410A can be made of glass and can be either hydrophilic or hydrophobic. When base assembly 1410A is glass or a polymeric film without an adhesive, the lower surface of mask assembly layer 1403B has a PCR-compatible adhesive thereon to facilitate bonding the base substrate layer assembly to the mask assembly layer after fingerprint nucleic acid deposition to form reaction wells for the in situ nucleic acid amplification reaction.

Another embodiment of the device is shown in Figures 17A and 17B. Device 1701A is shown in assembly and has a base substrate assembly 1710A made of a film compatible with both fingerprint collection and PCR reaction conditions. The upper surface area (including areas 1710A-1 and 1710A-2) film can be coated with a PCR-compatible adhesive. The fingerprint image and fingerprint nucleic acid 1702A are collected in area 1710A-1 when the device is placed on the scanning surface of the imaging assembly. Mask layer assembly 1703A with through hole 1707A is another part of the device. In this embodiment, ring assembly 1708A is bonded to mask assembly layer 1703A, surrounding the edge of through hole 1707A. Open PCR device 1701B is shown in its completed form in FIG17B and is formed by bonding a mask assembly layer 1703B, which can be a thin plastic or glass slide (e.g., 1 mm), having a 10 mm diameter through-hole 1707B, to a base substrate layer assembly 1710B containing a fingerprint nucleic acid 1702B still aligned with the opening of through-hole 1707B. Ring 1708B helps retain nucleic acid amplification reagents within the reaction well formed by the combination of base substrate layer 1710B and mask assembly layer 1703B. Open PCR can then be performed in situ on the fingerprint nucleic acid and sealed with a sealing solvent layer (including but not limited to an oil layer). Detection of the amplified nucleic acid in an aliquot can be performed by extracting the aliquot from under the sealing solvent layer.

In one variation of the device illustrated in Figures 17A-B, base substrate assembly 1710A is a sheet of transparent adhesive film (Life Technologies, 4306311). The open PCR device 1701B is formed by bonding a glass mask assembly layer 1703B having a 10 mm diameter through hole to the adhesive film of base substrate assembly layer 1710A containing the fingerprint nucleic acid deposited at region 1710A-1. In another variation, the upper surface of glass mask assembly layer 1703B is hydrophobic to help retain the sealing solution during the open PCR reaction. In another variation, base substrate assembly 1710A can be made of glass and can be either hydrophilic or hydrophobic. When base assembly 1710A is glass or a polymeric film without an adhesive, the lower surface of mask assembly layer 1703B has a PCR-compatible adhesive thereon to facilitate bonding the base substrate layer assembly to the mask assembly layer after fingerprint nucleic acid deposition to form reaction wells for the in situ nucleic acid amplification reaction.

Collection Aid Liquid. In various aspects of the present invention, when collecting a biological sample onto the base substrate assembly of the device, a collection aid liquid may be present. The collection aid liquid may be a solvent, a detergent, or a dissolving solution.

Solvents. Some suitable solvents that can help collect nucleic acids from the substrate can include water, ethanol, or acetonitrile. In some embodiments, the presence of one of these solvents can help collect more nucleic acids present in the fingerprint on the base substrate assembly.

As used herein, the term "detergent" is any substance that reduces the surface tension of water and is used synonymously with the term "surfactant". In certain embodiments, the detergent can be a cationic detergent, an anionic detergent, a nonionic detergent, or a zwitterionic detergent. Examples of nonionic detergents include tritons, such as the Triton ^{™ X series (Triton™ X} -100t-Oct-C ₆ H ₄ --(OCH ₂ --CH ₂ ) _x OH, x=9-10; Triton ^™ X-100R; Triton ^™ X-114x=7-8); octyl glucoside; polyoxyethylene (9) lauryl ether; digitonin; IGEPAL ^™ CA630 octylphenyl polyethylene glycol; n-octyl-β-D-glucopyranoside (βOG), n-dodecyl-β; Tween ^™ 20 polyethylene glycol sorbitan monolaurate; Tween ^™ 80 polyethylene glycol sorbitan monooleate; polydocanol; n-dodecyl β-D-maltoside (DDM); NP-40 nonylphenyl polyethylene glycol; C12E8 (octaethylene glycol n-dodecyl monoether); hexaethylene glycol mono-n-tetradecyl ether (C14EO6); octyl-β-thiopyranoside (octylthioglucoside, OTG); Emulgen; and polyoxyethylene 10 lauryl ether (C12E10). Examples of ionic detergents (anionic or cationic) include deoxycholate, sodium dodecyl sulfate (SDS), N-lauroyl sarcosine, and cetyltrimethylammonium bromide (CTAB). Zwitterionic agents can also be used in the purification process of the present invention, such as Chaps, zwitterion 3-14, and 3-[(3-cholamidopropyl)dimethylamino]-1-propanesulfonate. It is also contemplated that urea can be added with or without another detergent or surfactant.

In some embodiments, surfactants that are substantially free of fluorescence between 300 nm and 750 nm are used in the methods of the present invention. In some embodiments provided herein, when used for in situ analysis of DNA, RNA, protein, or surrogate thereof, the lysis solution includes a surfactant at a concentration that has low or no emission at the emission wavelength of a dye or label commonly used to detect RNA, DNA, or protein.

In some embodiments, the effective concentration of surfactant in the solubilization mixture is the concentration of surfactant at which the sample is considered completely solubilized as determined by propidium iodide staining using 1% TRITON X-100 ^™ surfactant as a control. Exemplary solubilization effective concentrations of surfactants range from 0.02% or 0.05% to 3% or more for TRITON X-114 ^™ surfactant, from 0.1% to 5% or more for NP-40 ^™ surfactant, and from 0.05% to 1% or to 3% for TRITON X-100 ^™ surfactant. When a combination of surfactants is used, the concentration of each surfactant can be reduced compared to the amounts recited.

The lysis solution may contain an enzyme that promotes nucleic acid collection onto the base substrate assembly. In certain embodiments, the lysis solution contains proteinase K. In various embodiments, proteinase K may be present in the lysis solution at about 0.8 mg/ml to about 1.5 mg/ml. In certain other embodiments, the lysis solution may contain a protease. The enzyme may degrade structural proteins to allow extraction of biomolecules from biological cells on the substrate. The enzyme may be applied to the substrate while the sample is being collected, or the enzyme may be incorporated into the base substrate assembly, for example, incorporated into a polymer film or coated on glass. In certain embodiments herein, the lysis solution includes a polypeptide with protease activity, for example proteinase K.

Instead of or in addition to proteinase K, the lysis solution can include a serine protease, such as trypsin, chymotrypsin, elastase, subtilisin, griseusin, thermolysin, hydrolysin, plasmin, cucumisin, or carboxypeptidase A, D, C, or Y; a cysteine protease, such as papain, calpain, or clostripain; an acid protease, such as pepsin, chymosin, or cathepsin; or a metalloprotease, such as pronase, thermolysin, collagenase, dispase, aminopeptidase, or carboxypeptidase A, B, E/H, M, T, or U. Qiagen protease (p/n 19155, Qiagen, Valencia, CA) is an alternative to proteinase K and may be inactivated by EDTA.

In other embodiments, the enzyme contained in the lysis solution is a trypsin hydrolase, for example porcine pancreatin. Keratinases that can be useful for nucleic acid collection include but are not limited to those isolated from bacteria or fungi. Some keratinase enzymes have increased stability in the presence of detergents, surfactants, metal ions, and solvents, which are suitable for the methods of the present invention. Non-limiting examples of keratinase enzymes suitable for use in the methods of the present invention include keratinase enzymes from Pichia pastoris, Bacillus megaterium, and Bacillus licheniformis.

Automated Operation. In some embodiments, a system providing at least one component of an apparatus can be operably loaded within a housing of the device. The apparatus is configured to automatically subject nucleic acids deposited on a base substrate assembly to a nucleic acid amplification reaction, which may further include a nucleic acid sequencing reaction. The housing may include a platform for securing at least the base substrate assembly containing the collected biological sample. After securing the substrate, the apparatus may initiate the nucleic acid amplification and/or sequencing reaction. The apparatus may further attach an identifier to at least one component of the apparatus. The identifier may be an identifier assigned to the substrate when the ridge and valley features and biological sample are collected, or it may contain additional information regarding the shipping, archiving, or handling of the substrate after collection. The apparatus may include a processor configured to control the apparatus to perform reactions and/or nucleic acid sequence identity detection. The processor may be configured to attach the identifier assigned to at least the base substrate assembly and may further be configured to attach any additional information regarding the shipping, archiving, or handling of the base substrate assembly after the reaction or detection process. The apparatus may also include computer-readable media to instruct the apparatus to perform concentration and identification operations. The computer-readable medium may be non-transitory.

Integrated Systems for Amplification and Identification. While microfluidics technology is commercially available for integrated nucleic acid amplification and identification, the relatively high cost (approximately $350 per sample) may make this approach unpopular given emerging improvements in direct amplification workflows for processing cheek swab samples. Microfluidic instruments for "Fast DNA" systems that integrate the functions of cell lysis, DNA purification, PCR, and capillary electrophoresis identification rely on on-chip microvalves and micropumps. The resulting complexity of the design of such microfluidic or dielectric cartridges increases the cost per sample.

Using recently commercially available STR analysis kits (e.g., Direct or Select (Life Technologies)), it has been found that neither extensive cell lysis nor nucleic acid purification is required to generate high-quality STR profiles directly from cheek swab samples. The cheek swab sample only needs to be placed in approximately 300 μl of buffer (Life Technologies) for approximately 10 minutes, and approximately 3 μl of the lysate can be transferred directly to the reaction site. Paper swabs from paper substrates can also be dissolved in a small volume of lysis buffer and amplified directly. The reaction site can be a hydrophilic location on a plate (see Figure 18) or a reaction well on a simple reaction cartridge (see Figure 21). A small amount (approximately 25 μl of STR PCR master mix) is added to the sample for nucleic acid amplification. Instead of a closed chamber or tube, PCR can be performed in an open PCR format, where a sealing solution (e.g., mineral oil) is used to prevent evaporation. Using open PCR, simple liquid dispensing techniques (including but not limited to the use of AutoMate Express instruments) can fully automate the STR analysis workflow at a lower cost, simplified handling, and workflow requirements.

Fingerprint nucleic acid collection and subsequent analysis can be automated using reaction plates (such as the one illustrated in FIG18 ). Open PCR slides contain hydrophilic wells defined by hydrophobic modification (e.g., printing) of the slide surface. The open PCR format allows for convenient recovery of PCR products through a sealing solution by simple pipetting. The small thermal cycler footprint and weight make it an ideal PCR module for fully integrated STR profiling. The AmpliSpeed slide thermal cycler used in Example 6 weighs only 2.9 kg and measures only approximately 126 mm × approximately 295 mm × approximately 112 mm.

Washing the fingerprint nucleic acid deposit and subsequently transferring the wash solution to an open PCR slide provides the option of using a wash solution that selectively dissolves nucleic acids and leaves PCR inhibitors from the fingerprint nucleic acid deposit on the substrate. The base substrate assembly surface 1810-1.1 (or occupied 1809) of Figure 18 can also be treated to have the same function of retaining inhibitors and releasing DNA into the wash solution.

Hybrid devices containing both fingerprint DNA collection and open PCR functionality improve chain-of-custody integrity because the fingerprint DNA never leaves the device before the nucleic acid amplification reaction is complete.

Due to the simplicity of the liquid transfer requirements ( FIG. 21 shows an exemplary cartridge schematic; the workflow is described below), the liquid handling arm only needs to transfer a volume of up to 25 μL and only needs to travel a distance of 10 cm in the XYZ direction. If this system is designed to process 8 samples simultaneously, the thermal cycler and CE module will be much smaller than current separate PCR and CE instruments. The fact that all three of the liquid handler, PCR, and CE components can be controlled by a single computer will also help reduce size and weight. The total run time can be shorter than commercially available microfluidic systems, such as the RapidHIT ^™ (Integenex), which requires ninety minutes. Because the cartridge is extremely simple and can be designed to run a single sample, the cost can be lower than that of the RapidHIT ^™ , which is configured to run four samples, which may incur losses.

Example workflow.

Place the cartridge into the instrument. The reservoir well containing the sealing solution resides within the PCR thermal cycler. The reservoir well containing formamide and GS500 size standards (Life Technologies) resides in a holder located on the translation stage, which allows transfer of formamide and size standards (Life Technologies) for CE injection after amplification.

• The cheek swab containing nucleic acid was placed into the wells containing approximately 300 μl (Life Technologies) lysis buffer.

● Close the instrument and start the experimental run.

• The instrument activates the cutter to cut the connection between the hole and the rest of the barrel.

• After the 10 minute incubation, the liquid dispensing arm picks up the pipette tip and transfers 3 μL of lysate into the PCR mix well containing the Direct Amplification STR Master Mix and agitates the mix several times.

The instrument transfers and places the PCR mixture containing nucleic acids into the bottom of a well containing a sealing solution (eg, mineral oil). The sealing solution will prevent the PCR reactants from evaporating.

PCR thermal cycling can be run as directed by the manufacturer's recommendations or the cycles can be modified as needed. A typical run might be 95°C/11 m, followed by 30 cycles of 94°C/20 s, 59°C/2 m, 72°C/1 m, followed by 60°C/25 min and a 4°C hold.

• Following PCR, transfer 1 μL of PCR product to the wells containing formamide and size standards.

• The wells containing formamide, size standards and amplified nucleic acids are transported via a translation stage to the CE module for CE analysis.

A system for identifying a target nucleic acid is provided, comprising: a base substrate assembly comprising one or more hydrophilic sites separated by hydrophobic regions, wherein at least one hydrophilic site is configured to contain at least one target nucleic acid, and further wherein the base substrate assembly is made of one or more materials compatible with the reaction conditions of a nucleic acid amplification reaction; a reaction assembly configured to amplify the at least one target nucleic acid at the at least one hydrophilic site to form at least one nucleic acid amplification product; and a detection assembly configured to identify at least one nucleic acid sequence of the at least one nucleic acid amplification product. In some embodiments, the base substrate assembly may be a polymeric film or glass. In various embodiments, the reaction assembly may not require a solid upper seal assembly for the substrate. The reaction assembly may be configured to provide an oil seal to the at least one hydrophilic site for conducting the nucleic acid amplification reaction. The reaction assembly may be configured to provide an oil seal to each of the one or more hydrophilic sites. The reaction assembly may be configured to provide nucleic acid amplification reagents to the at least one target nucleic acid contained on the at least one hydrophilic site on the base substrate assembly. The reaction component can be configured to provide suitable reaction conditions for nucleic acid amplification of the at least one target nucleic acid contained on the at least one hydrophilic site on the base substrate component. The system can be further configured to extract a volume of the at least one nucleic acid amplification reaction product from under the oil seal for use in identifying the at least one nucleic acid sequence using the detection component.

The detection component of the system can be selected from a fluorescent dye sequencing component, a semiconductor sequencing component, or a pyrophosphate sequencing component. In some embodiments, the detection component can be a fluorescent dye sequencing component. In some embodiments, the fluorescent dye sequencing component can be a mobility-dependent fluorescent dye sequencing component. In some embodiments, the mobility-dependent fluorescent dye sequencing component can be capillary electrophoresis.

The present invention may provide a base substrate assembly for nucleic acid amplification and identification, comprising: one or more hydrophilic sites separated by hydrophobic regions, wherein at least one hydrophilic site is configured to contain at least one target nucleic acid; wherein the base substrate assembly is made of one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction; and further wherein the base substrate assembly is configured to allow extraction of an aliquot of the amplified target nucleic acid for detection of the nucleic acid sequence identity. The base substrate assembly may be a polymer or glass.

A kit is provided, comprising the base substrate assembly as described above, and optionally instructions for use thereof. The kit may further comprise reagents for nucleic acid amplification reactions. The kit may further comprise reagents for nucleic acid sequencing reactions.

A method for identifying at least one target nucleic acid is provided, comprising the following steps: providing a base substrate assembly having one or more hydrophilic sites separated by hydrophobic regions, wherein at least one hydrophilic site comprises at least one target nucleic acid, and further wherein the base substrate assembly is made of one or more materials compatible with the reaction conditions of a nucleic acid amplification reaction; providing a reagent suitable for amplifying the at least one target nucleic acid at the at least one hydrophilic site; performing a nucleic acid amplification reaction on the at least one target nucleic acid to form at least one nucleic acid amplification product; and detecting the nucleic acid sequence of the at least one nucleic acid amplification product, thereby identifying the at least one target nucleic acid.

In some embodiments, a solid upper sealing component may not be provided for the substrate. In various embodiments, the method further comprises the steps of providing a sealing solution layer to the at least one hydrophilic site containing the at least one nucleic acid for the nucleic acid amplification reaction. In some embodiments, the sealing solution layer may be provided to each of the one or more hydrophilic sites. In some embodiments, the method further comprises the steps of providing a nucleic acid amplification reagent to the at least one target nucleic acid contained on the at least one hydrophilic site on the substrate. The method may further comprise providing suitable reaction conditions for performing nucleic acid amplification on the at least one target nucleic acid contained on the at least one hydrophilic site on the base substrate assembly. In some embodiments, a certain volume of the at least one nucleic acid amplification reaction product is extracted from under the oil seal to identify the at least one nucleic acid sequence by the detection component.

In some embodiments of the method, the detection component can be selected from a fluorescence detection component, a semiconductor detection component, or a pyrophosphate detection component. In other embodiments, the detection can be a fluorescent dye sequencing component. In other embodiments, the fluorescent dye sequencing component can be a mobility-dependent fluorescent dye sequencing component. In other embodiments, the mobility-dependent fluorescent dye sequencing component can be capillary electrophoresis.

The present invention also provides a system for identifying a target nucleic acid, comprising (a) a reaction cartridge, comprising: a dissolution well configured to accommodate a sample substrate having a biological sample, the sample substrate comprising the target nucleic acid and a dissolution buffer of an aliquot, wherein the biological sample is dissolved to release the target nucleic acid; a PCR well configured to accommodate a PCR reagent of an aliquot, wherein the released target nucleic acid is amplified to form an amplified target nucleic acid sample; a sealing solution well configured to accommodate a sealing solution of an aliquot, wherein the sealing solution is transferred to the PCR well to seal the open PCR; and optionally, a well configured to transfer the released A transfer end for any one of a target nucleic acid, a sealing solution, and an amplified target nucleic acid sample; and optionally, a detection sample preparation hole operably connected to the reaction barrel, wherein the detection sample preparation hole is configured to accommodate an aliquot of a sample configured to prepare a reagent for identifying the nucleic acid sequence of the amplified target nucleic acid sample; and wherein the reaction barrel comprises one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction; (b) a reaction component configured to amplify the released target nucleic acid to form the amplified target nucleic acid sample; and (c) a detection component configured to identify the nucleic acid sequence of the amplified target nucleic acid sample. In some embodiments, the reaction barrel can be made of a polymer or glass. In some embodiments, the sample substrate can be a length of paper substrate, including but not limited to a paper smear from a paper substrate on which a biological sample has been collected, or a cheek swab. In various embodiments, the reaction barrel may not require a solid upper closure component for the reaction barrel. In some embodiments, the system can be configured to transfer an aliquot of the released target nucleic acid from the dissolution hole to the PCR hole. In certain embodiments, the reaction assembly can be configured to provide suitable reaction conditions to carry out nucleic acid amplification to the target nucleic acid of the release contained in the PCR wells of the reaction barrel. In various embodiments, the system can be configured to extract a certain volume of the amplified target nucleic acid sample from under the oil seal and transfer it to the detection sample preparation hole to prepare the amplified target nucleic acid sample for identifying the nucleic acid sequence by the detection assembly. In other embodiments, the detection assembly can be selected from a fluorescent dye sequencing assembly, a semiconductor sequencing assembly or a pyrophosphate sequencing assembly. In certain embodiments, the detection assembly can be a fluorescent dye sequencing assembly. In certain embodiments, the fluorescent dye sequencing assembly can be a mobility-dependent fluorescent dye sequencing assembly. In certain embodiments, the mobility-dependent fluorescent dye sequencing assembly can be a capillary electrophoresis assembly. The system can further include a plurality of reaction barrels.

The present invention also provides a reaction tube for nucleic acid amplification and identification, comprising (a) a dissolution well configured to accommodate a sample substrate having a biological sample, the sample substrate comprising a dissolution buffer of the target nucleic acid and an aliquot, wherein the biological sample dissolves to release the target nucleic acid; (b) a PCR well configured to accommodate a PCR reagent of the aliquot, wherein the released target nucleic acid is amplified to form an amplified target nucleic acid sample; (c) a sealing solution well configured to accommodate a sealing solution of the aliquot, wherein the sealing solution is transferred to the PCR well to seal the open PCR; and optionally, (d) a transfer end configured to transfer any one of the released target nucleic acid, the sealing solution, and the amplified target nucleic acid sample of the aliquot; and optionally, (e) a detection sample preparation well operably connected to the reaction tube, wherein the detection sample preparation well is configured to accommodate aliquots of reagents configured to prepare the sample for identifying the nucleic acid sequence of the amplified target nucleic acid sample; and further wherein the reaction tube is made of one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction. In some embodiments, the reaction tube is made of one or more polymers, glass, or a combination thereof.

The present invention also provides a kit comprising a reaction tube for nucleic acid amplification and identification, the reaction tube comprising (a) a dissolution well configured to accommodate a sample substrate having a biological sample, the sample substrate comprising the target nucleic acid and a dissolution buffer of an aliquot, wherein the biological sample is dissolved to release the target nucleic acid; (b) a PCR well configured to accommodate an aliquot of PCR reagent, wherein the released target nucleic acid is amplified to form an amplified target nucleic acid sample; (c) a sealing solution well configured to accommodate an aliquot of sealing solution, wherein the sealing solution is transferred to the PCR well to seal the open PCR; and optionally, (d) a transfer end configured to transfer any one of the released target nucleic acid, the sealing solution and the amplified target nucleic acid sample of the aliquot; and optionally, (e) a detection sample preparation well operably connected to the reaction tube, wherein the detection sample preparation well is configured to accommodate an aliquot of reagent configured to prepare the sample for identifying the nucleic acid sequence of the amplified target nucleic acid sample; and further wherein the reaction tube is made of one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction, and optionally instructions for use thereof. In some embodiments, the reaction cartridges of the batch are made of one or more polymers, glass, or a combination thereof. In various embodiments, the test kit may further include reagents for nucleic acid amplification reactions. In various embodiments, the test kit may further include a sealing solution. In some other embodiments, the test kit may further include reagents for nucleic acid sequencing reactions.

The present invention also provides a method for identifying a target nucleic acid, which comprises the following steps: (a) providing a reaction cylinder, the reaction cylinder comprising: a dissolution well configured to accommodate a sample substrate containing a biological sample, the sample substrate comprising the target nucleic acid and a dissolution buffer of an aliquot, wherein the biological sample is dissolved to release the target nucleic acid; a PCR well configured to accommodate a PCR reagent of an aliquot, wherein the released target nucleic acid is amplified to form an amplified target nucleic acid sample; a sealing solution well configured to accommodate a sealing solution of an aliquot, wherein the sealing solution is transferred to the PCR well to seal the open PCR; and optionally, a transfer end configured to transfer any one of the released target nucleic acid, the sealing solution and the amplified target nucleic acid sample of the aliquot. part; and optionally, a detection sample preparation hole operably connected to the reaction barrel, wherein the detection sample preparation hole is configured to accommodate an aliquot of a reagent configured to prepare the sample for identifying the nucleic acid sequence of the amplified target nucleic acid sample; and wherein the reaction barrel includes one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction; (b) providing a reagent suitable for amplifying the released target nucleic acid in the PCR hole of the reaction barrel; (c) sealing the PCR hole with the sealing solution of the aliquot; (d) performing a nucleic acid amplification reaction on the released target nucleic acid to form the amplified target nucleic acid sample; and (e) detecting the nucleic acid sequence of the amplified target nucleic acid sample, thereby identifying the at least one target nucleic acid. In some embodiments, the reaction barrel can be a polymeric film or glass. In some embodiments, the sample substrate can be a section of paper substrate or a cheek swab. In various embodiments, the method can further include the steps of adding the sample substrate including the biological sample containing the target nucleic acid to the dissolution hole, and dissolving the biological sample to release the target nucleic acid. In other embodiments, the method may further include the step of transferring an aliquot of the released target nucleic acid from the dissolution well to the PCR well using the transfer tip after the biological sample has been dissolved to release the target nucleic acid. In various embodiments, a solid upper sealing assembly may not be provided for the substrate. In some embodiments, the method may further include the step of transferring the sealing solution using the transfer tip to the PCR well of the reaction cartridge containing the released nucleic acid and the aliquot of PCR reagent. In some embodiments, the method may further include the step of providing suitable reaction conditions to amplify the released target nucleic acid contained in the PCR well of the reaction cartridge to form the amplified target nucleic acid sample. In some embodiments, a volume of the amplified target nucleic acid sample may be extracted from beneath the oil seal using the transfer tip and transferred to the test sample preparation well. In other embodiments, the detection assembly may be selected from a fluorescent dye sequencing assembly, a semiconductor sequencing assembly, or a pyrophosphate sequencing assembly. In other embodiments, the detection assembly may be a fluorescent dye sequencing assembly. In some embodiments, the fluorescent dye sequencing assembly may be a mobility-dependent fluorescent dye sequencing assembly. In some embodiments, the mobility-dependent fluorescent dye sequencing assembly may be a capillary electrophoresis assembly. In some embodiments, the reagents configured to prepare the aliquot of the sample for identifying the nucleic acid sequence of the amplified target nucleic acid sample may include a denaturant. The denaturant may be formamide. In some embodiments, the reagents of the aliquot further include a size standard.

Additional Imaging Components. In another embodiment of the present teachings, the system may further include at least a second imaging component for collecting a second image of the individual. The second image of the individual may be the individual's face or a biometrically identifiable portion of the individual. Suitable biometric images that may be collected as the second image include retinal or iris scans, ear outlines, facial recognition, hand geometry, foot geometry, voice, smell, and fragrance.

Processor. After collecting at least one ridge and valley signature, the signature in analog or digital form can be transmitted to a database having a plurality of ridge and valley signatures and any other physical biometric data collected and deemed suitable for transmission. In some embodiments, the system includes a processor configured to transmit the ridge and valley signatures obtained from an individual to at least one database storing ridge and valley signatures for the individual. In some embodiments, the database is selected from the group consisting of: forensic, criminal, identity, child identity, missing persons, immigration, DMV, terrorist, paternity, arrestee, criminal database, access control, or any combination thereof.

After amplifying nucleic acids collected from an individual and detecting the nucleic acid sequence identity of the amplicons produced by the amplification, one or more processors of the system can be configured to transmit the nucleic acid sequence identity of the individual to at least one database containing nucleic acid sequence identities, the database being selected from the group consisting of: forensic, criminal, identity, child identity, missing persons, immigration, DMV, terrorist, paternity testing, arrestee, criminal database, access control, and any combination thereof.

Identifier. In other embodiments of the present teachings, the system may further include an identifier for associating identification information with an individual's ridge and valley signatures, a biological sample containing nucleic acid, and optionally a physical image (including any of a facial image, an iris image, a retinal image, or an ear image). In some embodiments, the individual's name is included in the identifier. The identifier can help associate various collected samples and eliminate sample confusion and human error that can occur with non-systematic sample labeling used to identify collected data and samples. In various embodiments, the identifier is alphanumeric, graphical, magnetic, or electromagnetic. In some embodiments, the identifier is a barcode. In various embodiments, the base substrate assembly includes the identifier. In other embodiments, the frame requires the identifier. In some embodiments, the packaging for the device includes the identifier. In other embodiments, both the substrate assembly and the packaging include identifiers to help associate collected samples and images. If more than one imaging assembly is included in the system, the same identifier can be used for all collected data and samples for an individual.

Reaction Assembly. In other embodiments, the system further includes an amplification assembly, which can be used to optionally release at least one nucleic acid from a biological sample; amplify at least one nucleic acid; and/or isolate the amplified at least one nucleic acid. The reaction assembly is configured to subject a device containing a biological sample on a substrate to a nucleic acid amplification reaction, which can include thermal cycling conditions. This includes any suitable thermal cycling instrument that can be used or adapted to contain the devices described herein. The reaction assembly can be configured to subject a single biological sample containing nucleic acid, i.e., a device having only a single area for a single fingerprint nucleic acid deposition. Alternatively, the reaction assembly can be configured to subject multiple biological samples containing nucleic acid to a nucleic acid amplification reaction, such as a single sample from multiple individuals as shown in Figure 2, or multiple devices having a single fingerprint nucleic acid deposition site. When the reaction assembly is configured to subject multiple devices configured to contain a single fingerprint nucleic acid deposition site, the reaction assembly can include a frame to secure the multiple devices within the reaction assembly instrument for safe disposal. The nucleic acid amplification reaction can be a polymerase chain reaction, and can be performed in an open PCR format. The reaction assembly can further be configured to perform a DNA sequencing reaction while the nucleic acid and/or nucleic acid amplification products are still present on the substrate assembly of the device. The nucleic acid sequencing reaction can be a fluorescent dye sequencing reaction, a semiconductor sequencing reaction or a pyrophosphate sequencing reaction. When the reaction is a fluorescent sequencing reaction, the fluorescent sequencing reaction can be sequencing by synthesis or Sanger sequencing (Sanger sequencing). The biological sample containing at least one nucleic acid can be subjected to analysis and DNA sequencing methods for nucleic acid markers (e.g., DNA markers for STRs, Indels, SNPs and combinations thereof). The reagents used to analyze nucleic acids are commercially available, such as direct PCR amplification kits or Select Express (Applied Biosystems, Foster City, California) and 18D systems (Promega Corp., Madison, Wisconsin, Madison, WI) according to the manufacturer's instructions.

The reaction assembly can be configured to perform nucleic acid amplification reactions and/or sequencing reactions in an open PCR format, i.e., wherein the reaction wells formed in the device are sealed with a layer of sealing solution to prevent evaporation during the reaction. In some other embodiments, the reaction assembly is configured to perform nucleic acid amplification reactions and/or sequencing reactions when the device is sealed with a cover assembly.

Sealing solution. In embodiments utilizing open PCR, the nucleic acid amplification and/or sequencing reaction mixture in the reaction wells of the device is protected from evaporation or contamination by a sealing solution layer on the reaction mixture. The sealing solution can be any suitable fluid or semi-fluid that is immiscible or forms an immiscible layer on the reaction mixture. The sealing solution can be a synthetic substance or a natural product. In some embodiments, the density of the sealing solution can be less than the density of the reaction mixture sealed by the sealing solution. In some embodiments, the sealing solution may not inhibit the nucleic acid amplification reaction and/or sequencing reaction. The sealing solution can be stable at both high and low temperatures. The sealing solution may not be mixed into the nucleic acid amplification mixture and/or sequencing reaction mixture. In some embodiments, the sealing solution is oil. In some embodiments, the sealing solution can be mineral oil.

Detection assembly. The system can include a detection assembly, wherein the detection assembly is configured to detect at least one product of nucleic acid amplification. This detection can identify the corresponding nucleic acid sequence of the nucleic acid amplification product, which can provide individual nucleic acid sequence identification. The detection assembly can be a fluorescent dye detection assembly, a semiconductor detection assembly, or a pyrophosphate detection assembly. In some embodiments, the detection assembly is a fluorescent dye detection assembly, which can optionally be a mobility-dependent fluorescent dye detection assembly. In some embodiments, the mobility-dependent fluorescent dye detection assembly is used with a capillary electrophoresis instrument with fluorescence detection.

Any or all of these additional components may be used to identify an individual.

Kit. The present invention also provides a kit. The kit may contain one or more devices of any of the various embodiments of the device described herein, wherein the device may have any combination of features as described herein, and may optionally contain instructions for use of the device. The kit may further include a collection auxiliary liquid, including but not limited to a solvent, a detergent, or a dissolving solution. The kit may further include reagents for stabilizing biological samples on the base substrate assembly of the device for archiving or shipping. The kit may further include reagents for analyzing samples, including but not limited to antibodies, stains, indicators, agar plates, or nucleic acid amplification reagents. The kit may contain other reagents for reaction (e.g., PCR amplification reaction, nucleic acid sequencing reaction) or analysis (e.g., STR, SNP, or Indel analysis). The kit may contain a sealing solution for sealing the reaction mixture on the base substrate assembly during the reaction. The kit may contain one or more proteases for extracting nucleic acids from biological samples collected to the base substrate assembly of the device.

The kit may include storage packaging for the device containing the biological sample and/or nucleic acid and may also include archiving instructions. The kit may further include mailing packaging for the device containing the biological sample or nucleic acid.

The packaging included in the test kit can include a frame to prevent contamination of any part of the device. Any part of at least one package can include an identifier to associate any part of the device with the ridge and valley characteristic marks obtained when collecting the biological sample. The identifier can further associate the device, the base substrate assembly of the device, the ridge and valley characteristic marks with the individual providing the sample in any combination. In various embodiments, the identifier is alphanumeric, graphical, magnetic, or electromagnetic. In some embodiments, the identifier is a bar code.

In some embodiments, the kit may include instructions for use of the device; storage of the substrate or device after sample collection; performing nucleic acid amplification and/or nucleic acid sequencing reactions on the substrate of the device; and transferring an image of at least one ridge and valley feature to a database. The kit may also include instructions for mailing the device to another facility for analysis or archiving after sample collection.

Method. Both fingerprints and biological samples of sufficient quality/quantity can be provided to identify an individual. An image of a fingerprint obtained by scanning through a base substrate assembly and optionally a mask assembly can be provided in high quality. When the image of the fingerprint is processed according to the requirements of any organization that has a database for reference, the digitized fingerprint can meet threshold requirements. As can be seen in Examples 2 and 4, and particularly in Figures 5 and 11, fingerprints obtained according to the method described herein can be obtained that are scored as the highest level NIST quality score of 1. Thus, a high-quality fingerprint image is provided, which can be electronically processed in any suitable manner to specify feature points or other classification features and is suitable for transmission to any organization for database comparison or for storage in a database. A non-limiting example is the Integrated Automated Fingerprint Identification System (IAFIS) maintained by the Federal Bureau of Investigation (a U.S. government agency).

In some embodiments, the at least first imaging component is an optical scanner, wherein the optical scanner includes an LED, a laser diode, an incandescent light source, or a multispectral imager. The at least first imaging component may alternatively be a camera, an active pixel imager, a CMOS imager, an imager that images in multiple wavelengths, a CCD camera, a photodetector array, and a TFT imager. The at least one ridge and valley feature can be collected electronically. The scanning surface of the at least first imaging component can be formed of a material selected from the group consisting of plastic, polyolefin, polystyrene, metal, metal alloy, glass, silicon, or a combination thereof. In some embodiments, the material of the scanning surface is transparent or translucent.

In order to obtain nucleic acid of sufficient quantity and quality from a fingerprint, several factors may need to be addressed. To ensure sufficient quantity, the individual may be instructed to touch a portion of their face once or several times with the finger to be imaged, and then firmly press the finger onto a base substrate assembly placed on the scanning portion of the fingerprint imaging system. In some embodiments of the method, this may not be necessary. The base substrate assembly suitable for use in the method may be a polymeric film or glass. In some embodiments, the polymeric film base substrate assembly may be a synthetic polymeric film. In other embodiments, the polymeric film base substrate assembly may be a natural polymer, including but not limited to starch, agarose, alginate, carrageenan, etc. In some embodiments, the base substrate assembly may be non-adhesive. In various embodiments, the base substrate assembly may be transparent or translucent.

Another aspect is obtaining high-quality STR profiles from nucleic acids deposited simultaneously with the fingerprint. In some cases, concurrent effects may arise from the presence of other substances in the deposited biological sample. While commercially available direct amplification STR chemistries are optimized for biological samples that may contain indigo, hemoglobin, and humic acid (all inhibitors of the PCR amplification process), no system is optimized for the types of inhibitors that may be present in biological samples obtained from fingerprint deposits. Depending on the specific donor and the physiological condition of the individual's skin at the time of fingerprinting, varying amounts of sebum and sweat may be present. Sebum is produced by sebaceous glands and contains wax monoesters (approximately 25%), triglycerides (approximately 40%), free fatty acids (approximately 15%), and squalene (approximately 10-15%). Sweat includes salts, urea, sugars, and ammonia. Furthermore, substances such as cosmetics, hair products, and sunscreens may be present when an individual deposits a fingerprint image containing a biological sample. Some or all of these chemicals may contribute to PCR inhibition. The normal skin pH of approximately 5.4 may also affect PCR efficiency under direct amplification workflows. Adjustment of the PCR master mix composition can reduce the inhibitory effects of any of these substances. A variety of master mixes/buffers, commercially available buffers, and other combinations were evaluated, and it was found that the Global master mix provided the most consistent results. This particular master mix provides higher concentrations of one or more polymerases and an increased percentage of bovine serum albumin (BSA). Both of these master mix components help overcome inhibitors found in DNA samples obtained from fingerprints. It has been surprisingly discovered that the use of a sample concentration wipe to concentrate the sample can be combined with direct amplification as described herein to provide high-quality STR analysis from highly limited amounts of nucleic acid deposited within a fingerprint, even in the presence of interference from other substances. As can be seen in Examples 1-5 and accompanying Figures 3, 6, 9, 12, 15, and 16, electropherogram analysis can conclusively identify the individual who donated the nucleic acid.

A method for collecting a biological sample containing at least one nucleic acid and collecting at least one ridge and valley characteristic of an appendage of an individual is provided, comprising the steps of providing at least a first imaging component. The at least first imaging component is configured to provide an energy wave capable of imaging the at least one ridge and valley characteristic; and has a scanning surface configured to allow the energy wave to penetrate the scanning surface. The method further comprises the steps of providing an apparatus having a base substrate component having at least a first surface area, wherein the base substrate component is configured to allow the energy wave to penetrate the substrate; collecting the biological sample from the appendage of the individual; and being made of one or more materials configured to be compatible with the reaction conditions of a nucleic acid amplification reaction. The apparatus can be any of the embodiments of the apparatus described herein having any combination of components. The method further comprises the steps of placing the appendage of the individual on the scanning surface and in contact with the base substrate component, thereby collecting the biological sample; and collecting the at least one ridge and valley characteristic from the appendage imaged by the energy wave. The appendage imaged can be a finger, a toe, a palm, or a sole of a foot. The steps of collecting the biological sample and collecting the at least one ridge and valley characteristic can be performed simultaneously.

In some embodiments, the at least first imaging component may be an optical scanner or a capacitive scanner, and the at least one ridge and valley feature may be collected electronically.

The method may further include the steps of transmitting the at least one ridge and valley signature of the individual to at least one database containing ridge and valley signatures, the database selected from the group consisting of: forensic, criminal, identity, child identity, missing persons, immigration, motor vehicle department, terrorist, paternity testing, arrestee, criminal database, access control and any combination thereof.

The method may further include the step of providing an identifier on at least one component of the device to associate the biological sample of the individual with the at least one ridge and valley characteristic of the individual, wherein the identifier is alphanumeric, graphical, magnetic, or electromagnetic. In some embodiments, the identifier is a barcode.

The method may further comprise the step of archiving at least the base substrate assembly of the device after performing the step of collecting the biological sample.

The method may further comprise the step of mailing the device to another location for archiving or testing after performing the step of collecting the biological sample.

The method may further require the step of subjecting the biological sample collected on the base substrate assembly to a nucleic acid amplification reaction. In some embodiments, the nucleic acid amplification reaction is a polymerase chain reaction. The method may further require the step of performing a nucleic acid sequencing reaction while the biological sample is still present on the base substrate assembly. In some embodiments, the nucleic acid amplification reaction and the nucleic acid sequencing reaction can be performed in the same reaction well without transferring or stopping between processes. In some embodiments, the amplification reaction can be a component of an STR analysis, a SNP analysis, or an Indel analysis.

The method may further require the step of placing a mask assembly of the device on the base substrate assembly, thereby surrounding at least the first surface area of the base substrate assembly. The mask assembly may be made of one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction. In some embodiments, the step of placing the mask assembly on the base substrate assembly is performed before the step of collecting the biological sample. In other embodiments, the step of placing the mask assembly on the base substrate assembly is performed after the step of collecting the biological sample and before the step of subjecting the biological sample on the base substrate assembly to the nucleic acid amplification reaction. The method may further include the step of sealing the nucleic acid amplification reaction with a sealing solution. The reaction well may not have any other upper closure besides the sealing solution. Alternatively, the method may include the step of placing a cover assembly of the device on the biological sample collected on the base substrate assembly during the nucleic acid amplification reaction. In some embodiments, the step of placing the cover assembly on the biological sample on the base substrate assembly is performed after the reagents for the nucleic acid amplification reaction have been added to the biological sample on the base substrate assembly and before the reaction begins. The method may further include the step of placing a cover assembly on the biological sample collected on the base substrate assembly for shipping or archiving prior to nucleic acid amplification. The method may further include the step of detecting the sequence of the nucleic acid, thereby identifying the individual. The method may further include the step of transmitting the nucleic acid sequence identity of the individual to at least one database containing nucleic acid sequence identities, the database selected from the group consisting of: forensic, criminal, identity, child identity, missing persons, immigration, Department of Motor Vehicles, terrorist, paternity, arrestee, criminal database, access control, and any combination thereof.

A method for identifying an individual is provided, comprising the steps of providing at least a first imaging component configured to provide an energy wave capable of imaging at least one ridge and valley feature of an appendage of the individual; and comprising a scanning surface configured to allow the energy wave to penetrate the scanning surface. The method further comprises the steps of providing an apparatus comprising a base substrate assembly having at least a first surface area, wherein the substrate assembly is configured to allow the energy wave to penetrate the substrate; collecting a biological sample comprising nucleic acid from the appendage of the individual; and being made of one or more materials configured to be compatible with reaction conditions of a nucleic acid amplification reaction. The apparatus can be any of the embodiments of the apparatus described herein having any combination of components. The method further comprises the steps of positioning the appendage of the individual on the scanning surface and in contact with the at least first surface area of the base substrate assembly, thereby collecting the biological sample; collecting the at least one ridge and valley feature from the appendage imaged by the energy wave, thereby providing identification of the ridge and valley feature of the individual; subjecting the nucleic acid of the biological sample collected on the base substrate assembly to a nucleic acid amplification reaction; and detecting the sequence of the nucleic acid, thereby identifying the individual. The steps of collecting the biological sample and collecting the at least one ridge and valley feature may be performed simultaneously.

In some embodiments, the at least first imaging component can be an optical scanner or a capacitive scanner. In various embodiments, the at least one ridge and valley feature can be collected electronically. In some embodiments, the appendage can be a finger, toe, palm, or sole. In other embodiments, the nucleic acid amplification reaction can be a polymerase chain reaction. In various embodiments, the method can include the step of performing a DNA sequencing reaction while still present on the base substrate assembly. In some embodiments, the amplification reaction can be a component of an STR analysis, a SNP analysis, or an Indel analysis.

The method may further include placing a mask assembly of the device on the base substrate assembly so as to surround at least the first surface area of the substrate, wherein the mask assembly is made of one or more materials compatible with the reaction conditions of a nucleic acid amplification reaction. In some embodiments, placing the mask assembly on the base substrate assembly may be performed before collecting the biological sample. In other embodiments, placing the mask assembly on the base substrate assembly may be performed after collecting the biological sample and before subjecting the biological sample on the substrate assembly to a nucleic acid amplification reaction.

The method may further comprise the step of sealing the nucleic acid amplification reaction mixture with a sealing solution.The nucleic acid amplification reaction mixture may not have other upper packaging.

The method may further comprise the step of placing a cover assembly of the device over the biological sample collected on the base substrate assembly. The cover assembly may be placed over the at least first surface of the base substrate assembly containing the nucleic acid after the biological sample is collected to facilitate shipping and archiving the device. In some embodiments, the cover assembly may be placed over the biological sample on the substrate assembly after reagents for the nucleic acid amplification reaction have been added to the biological sample on the base substrate assembly and before the reaction begins.

The method may further include the steps of transmitting the at least one ridge and valley signature of the individual to at least one database containing ridge and valley signatures, the database selected from the group consisting of: forensic, criminal, identity, child identity, missing persons, immigration, motor vehicle department, terrorist, paternity testing, arrestee, criminal database, access control and any combination thereof.

The method may further include the step of providing an identifier on at least one component of the device to associate the biological sample of the individual with the at least one ridge and valley characteristic of the individual, wherein the identifier is alphanumeric, graphical, magnetic, or electromagnetic. In some embodiments, the identifier is a barcode.

The method may further comprise the step of archiving at least the base substrate assembly of the device after performing the step of collecting the biological sample.

The method may further comprise the step of mailing the device to another location for archiving or testing after performing the step of collecting the biological sample.

The method may further comprise the step of transmitting the nucleic acid sequence identity of the individual to at least one database comprising nucleic acid sequence identities, the database being selected from the group consisting of forensic, criminal, identity, child identity, missing persons, immigration, DMV, terrorist, paternity, arrestee, criminal database, access control, and any combination thereof.

Those skilled in the art will appreciate that many modifications, alternatives, and equivalents are possible. All such modifications, alternatives, and equivalents are intended to be encompassed herein.

Examples

The following procedures are representative of procedures that may be used to collect, analyze, and archive/catalog biological samples and biometric data from individuals.

Example 1.

Use device as shown in Figure 2.By making one or more fingers of at least one individual be placed on the base substrate assembly made on the scanning surface of Lumidign ^TM optical scanner by a piece of about 2.5cm × about 5cm transparent adhesive film, make fingerprint imaging.By being placed on the adhesive film top containing fingerprint nucleic acid sediment by SecureSeal 8 hole gaskets (Grace Bio-labs, AB8R-0.5), open PCR reaction chamber (diameter is 8mm) (Fig. 2) is formed on each fingerprint.In this example, two chambers (210-1.3 and 210-1.4) on the upper left contain 007 control DNA, two chambers (210-1.1 and 210-1.2) on the lower left are formed on the fingerprint from donor 1, and four chambers (210-1.5, 210-1.6, 210-1.7 and 210-1.8) on the right are on the fingerprint from donor 2. Each chamber on the fingerprint was filled with 50 μl of Select Express PCR reaction mixture, which includes primers, polymerase, dNTPs, dye-labeled ddNTPs, and other reagents and buffers for nucleic acid amplification. Two positive control chambers were filled with 50 μl of Select Express PCR reaction mixture containing 5 ng of 007 control DNA. The top of the chamber was sealed with another piece of transparent adhesive film. The assembled fingerprint PCR device was placed on the AB9700 flat thermal cycler block and secured to the thermal block with tape.

The novel properties of the device in Figure 2 require modification of typical thermal cycling conditions to achieve optimal results. The resulting modified PCR thermal cycling conditions are: 96.5°C/1:30m, (98°C/15s, 56°C/46sec, 66°C/49sec) 29 times, 60°C/7min, 4°C-hold.

After thermal cycling, a 1 μl aliquot of each PCR reaction was mixed with a GS500 size standard and deionized formamide and analyzed using an ABI 3130xl capillary electrophoresis instrument using the default conditions for Select Express PCR samples: oven: 60°C, pre-run: 15 kV for 180 s, injection: 3 kV for 10 s, run: 15 kV for 1500 s, capillary length: 36 cm, separation polymer: POP4 ^™ polymer with dye set: G5. The resulting STR electropherograms were analyzed using ID-X software (Applied Biosystems). A complete STR profile for all 16 STR markers and the sex determination marker amelogenin was obtained for samples from one of reaction wells 210-1.5, 210-1.6, 210-1.7, or 210-1.8 (Figure 3). From left to right in the top lane, alleles for D10S1248, vWA, D16S539, and D2S1338 are shown; the second lane from the top of the page shows alleles for amelogenin, D8S1179, D21S11, and D18S51; the third lane from the top shows alleles for D22S1045, D19S433, TH01, and FGA; and the bottom panel shows alleles for D2S441, D3S1358, D1S1656, D12S391, and SE33.

Example 2.

Use the device as shown in Figure 4 and place it on the scanning surface of the Lumidign ^™ optical scanner. Cover assembly 404 is flipped open. By making the individual's finger contact and cover the entire area 410-1, fingerprint imaging is performed. The fingerprint image obtained (Figure 5) has a NIST quality score of 1. The cover assembly can be flipped closed so that it can be mailed to a central laboratory or it can be processed after being removed from the fingerprint scanning assembly. The cover assembly layer and the mask assembly layer can be removed before the nucleic acid amplification deposited in area 410-1. The base substrate assembly layer is transferred to a thermal cycler (including but not limited to AB9700 flat thermal cycler block) and fixed on the heat block.

The assembled fingerprint PCR device was placed on an AB 9700 flat thermal cycler block and secured to the thermal block with tape. Select Express PCR reaction mixture (50 μl) was added to area 410-1 containing nucleic acids collected by contact of the finger with the base substrate assembly. The PCR reaction mixture may contain a PCR-compatible colored dye to aid in subsequent PCR product recovery.

The PCR reaction mixture is sealed by adding a sealing solution (e.g., mineral oil) to the PCR reaction mixture. The novel nature of the open PCR device of Figure 4 requires modification of typical thermal cycling conditions to achieve optimal results. The resulting modified PCR thermal cycling conditions are: 96.5°C/1:30 min, (98°C/15 s, 56°C/46 sec, 66°C/49 sec) 29 times, 60°C/7 min, 4°C hold.

After thermal cycling, a 1 μl aliquot of each PCR reaction was mixed with a GS500 size standard and deionized formamide and analyzed using an ABI 3130xl capillary electrophoresis instrument using the default conditions for Select Express PCR samples: oven: 60°C, pre-run: 15 kV, 180 s, injection: 3 kV, 10 s, run: 15 kV, 1500 s, capillary length: 36 cm, separation polymer: POP4 ^™ polymer and dye set: G5. The resulting STR electropherogram was analyzed using ID-X software (Applied Biosystems). A complete STR profile was obtained for all 16 STR markers and the sex determination marker amelogenin ( FIG6 ). From left to right in the top lane, alleles for D10S1248, vWA, D16S539, and D2S1338 are shown; the second lane from the top of the page shows alleles for amelogenin, D8S1179, D21S11, and D18S51; the third lane from the top shows alleles for D22S1045, D19S433, TH01, and FGA; and the bottom panel shows alleles for D2S441, D3S1358, D1S1656, D12S391, and SE33.

The addition of PCR reagents and the addition of sealing solution do not have to be performed in the order described above. Similarly, a sealing solution of mineral oil can be added to the area 410-1 containing the fingerprint nucleic acid deposit on the base substrate assembly, and then 50 μl can be pipetted through the sealing solution layer to contact the fingerprint nucleic acid deposit in area 410-1.

Example 3.

Device 810A (Figure 8) further demonstrates the feasibility of capturing DNA from the fingertips on a PCR-compatible substrate and then forming a PCR reaction chamber directly on the fingerprint on the substrate for in-situ direct STR analysis. The device is placed on the scanning surface of a Lumidign ^™ optical scanner to collect fingerprint images and nucleic acid deposits by contact of a finger on the base substrate assembly 810A. In one form of this device, fingerprint nucleic acids are collected on a base assembly substrate 810A made of a transparent adhesive film (Life Technologies, 4306311). By placing a SecureSeal well (Grace BioLabs, AB8R-0.5) on the base assembly layer adhesive film 810A surrounding the area 810A-1 containing the fingerprint nucleic acid deposit 802A, a reaction chamber (about 8 mm in diameter and about 1 mm in depth) is formed on the area 810A-1 where the biological sample containing the fingerprint nucleic acid 802A is deposited. After bonding the two layers, the resulting chamber is filled with 50 μl of Select Express PCR reaction mixture. The top of the chamber is sealed with another blank transparent adhesive film. The assembled fingerprint PCR device was placed on an AB 9700 flat thermal cycler block and secured. The thermal cycling conditions were: 96.5°C/1:30 min, 29 cycles of (98°C/15 s, 56°C/46 sec, 66°C/49 sec), 60°C/7 min, and a 4°C hold. After thermal cycling, the top cover of the PCR chamber was cut open with a knife. The PCR reaction was recovered using a pipette and mixed with a GS500 size standard and deionized formamide. The reaction was analyzed using an AB 3130xl capillary electrophoresis instrument using the following conditions: oven: 60°C, pre-run: 15 kV, 180 s, injection: 3 kV, 10 s, run: 15 kV, 1500 s, capillary length: 36 cm, separation polymer: POP4 ^™ polymer and dye set: G5. The resulting STR electropherograms were analyzed using ID-X software (Applied Biosystems). A complete STR profile was obtained for all 16 STR markers and the sex determination marker amelogenin (Figure 9). The STR profile quality can be further improved by further optimizing the PCR thermal cycling conditions. In the top lane, from left to right, alleles for D10S1248, vWA, D16S539, and D2S1338 are shown; the second lane from the top of the page shows alleles for amelogenin, D8S1179, D21S11, and D18S51; the third lane from the top shows alleles for D22S1045, D19S433, TH01, and FGA; and the bottom panel shows alleles for D2S441, D3S1358, D1S1656, D12S391, and SE33.

Example 4.

In one embodiment, the apparatus 1001 ( FIG. 10 ) may be manufactured using the following procedure:

1. Place a piece of 11 mm diameter adhesive tape on the bottom of the concave depression on the glass base substrate assembly 1010.

2. Apply a few drops of a commercially available silanizing agent (eg, Silane®) to the glass base substrate assembly 1010. Use a cotton swab to wipe the remaining exposed glass surface to remove the availability of hydroxyl groups on the wiped portion of the glass base substrate assembly 1010.

3. Repeat step 2 once.

4. Clean the slide surface with a paper towel.

5. Remove the sticky tape.

6. Place a transparent mask assembly 1003 made of polymeric film with approximately 10 mm diameter holes on the slide, aligning the holes in the mask assembly with the untreated hydrophilic areas on the slide.

During fingerprint collection, the device 1001 was placed on a Lumidigm fingerprint scanner to acquire a fingerprint image of the index finger in contact with the hydrophilic area 1010-1. A high-quality fingerprint image with a NIST quality score of 1 was obtained (Figure 11). During STRPCR analysis, the transparent mask assembly 1003 was removed from the device 1001. The device 1001 containing the fingerprint nucleic acid deposit in the area 1010-1 was placed on an AB 9700 thermal cycler. 10 μl of Select Express PCR mixture was added to the fingerprint nucleic acid deposited in the hydrophilic area 1010-1. The reaction mixture was sealed with a layer of mineral oil (approximately 200 μl). The thermal cycling conditions were: 96.5°C/1:30m, 29 cycles of (98°C/15s, 56°C/46sec, 66°C/49sec), 60°C/7min and 4°C-hold. After thermal cycling, the PCR reaction was recovered using a pipette and mixed with a GS500 size standard and deionized formamide. An aliquot of this mixture was analyzed using an AB 3130xl capillary electrophoresis instrument using the following conditions: oven: 60°C, pre-run: 15 kV for 180 s, injection: 3 kV for 10 s, run: 15 kV for 1500 s, capillary length: 36 cm, separation polymer: POP4 ^™ polymer and dye set: G5. The resulting STR electropherogram was analyzed using ID-X software (Applied Biosystems). A complete STR profile was obtained for all 16 STR markers and the sex determination marker amelogenin ( FIG12 ). From left to right in the top lane, alleles for D10S1248, vWA, D16S539, and D2S1338 are shown; the second lane from the top of the page shows alleles for amelogenin, D8S1179, D21S11, and D18S51; the third lane from the top shows alleles for D22S1045, D19S433, TH01, and FGA; and the bottom panel shows alleles for D2S441, D3S1358, D1S1656, D12S391, and SE33.

Example 5.

The first example device 1401A ( FIG. 14A ) has a base substrate assembly 1410A made of a film compatible with both fingerprint collection and PCR reaction conditions. Base substrate assembly 1410A is coated with a PCR-compatible adhesive. A fingerprint image is collected and fingerprint nucleic acid 1402A is deposited within region 1410A-1. When a mask assembly 1403A, made of a thin (e.g., 1 mm thick) glass slide with through-holes 1407A approximately 10 mm in diameter, is bonded to base substrate 1410A containing fingerprint nucleic acid in region 1410A-1, an open PCR reaction well is formed. The through-holes are aligned to maintain access to region 1410A-1. Once bonded, the open PCR device ( 1401B) is ready for in situ amplification of the fingerprint nucleic acid deposit 1402B without requiring any handling or transfer.

The second example device 1403B is formed in the same manner as described in the previous paragraph, except that the upper surface of the mask assembly 1403B is treated to produce a hydrophobic surface, such as by treatment with a silanization agent, so as to retain reagents used in the nucleic acid amplification reaction.

Both the first and second apparatus 1403B were on an AB 9700 thermal cycler. PCR master mix (20 μl Select Expess PCR mix) was added to the exposed base substrate assembly area containing the fingerprint nucleic acid and the mix was covered with approximately 200 μl mineral oil to seal the reaction for open PCR.

Thermal cycling conditions were: 96.5°C/1:30 min, 29 cycles of 98°C/15 s, 56°C/46 sec, 66°C/49 sec, 60°C/7 min, and 4°C hold. Following thermal cycling, aliquots of each PCR reaction were recovered using a pipette and individually mixed with a GS500 size standard and deionized formamide. Each individual sample was analyzed using an AB 3130xl capillary electrophoresis instrument using the following conditions: oven: 60°C, pre-run: 15 kV, 180 s, injection: 3 kV, 10 s, run: 15 kV, 1500 s, capillary length: 36 cm, separation polymer: POP4 ^™ polymer with dye set: G5. The resulting STR electropherograms were analyzed using ID-X software (Applied Biosystems). Complete STR profiles were obtained for all 16 STR markers and the sex determination marker amelogenin for both open PCR devices. See Figure 15 (a first device having a hydrophilic upper surface of the mask assembly) and Figure 16 (a second device having a hydrophobic surface of the mask assembly). In the top lane, from left to right, alleles for D10S1248, vWA, D16S539, and D2S1338 are shown; the second lane from the top of the page shows alleles for amelogenin, D8S1179, D21S11, and D18S51; the third lane from the top shows alleles for D22S1045, D19S433, TH01, and FGA; and the bottom panel shows alleles for D2S441, D3S1358, D1S1656, D12S391, and SE33.

Example 6.

Biological samples containing nucleic acids were deposited by finger contact and collected on 3M PP2200 membranes in triplicate. The deposits were then rinsed with 25 μl of Select Express PCR reaction mixture. Each of three 5 μl rinses was transferred to a separate well of a Teflon-printed slide with a 4 mm pore diameter. The rinses were then covered with a drop of mineral oil. Three wells, each containing 5 μl of Select Express PCR reaction mixture plus 1 ng of 007 control DNA, were also loaded onto the same slide. The slide was then placed in a thermal cycler and the PCR reaction was performed using the following thermal cycling conditions: 95°C/1 min, (94°C/3 s, 59°C/16 sec, 65°C/29 sec) 29 times, 60°C/5 min, 4°C hold.

After thermal cycling, PCR reactions were recovered individually and mixed with a GS500 size standard and deionized formamide. Each individual sample was analyzed using an AB 3130xl capillary electrophoresis instrument using the following conditions: oven: 60°C, pre-run: 15 kV, 180 s, injection: 3 kV, 10 s, run: 15 kV, 1500 s, capillary length: 36 cm, separation polymer: POP4 ^™ polymer and dye set: G5. The resulting STR electropherograms were analyzed using ID-X software (Applied Biosystems).

Of the three fingerprint wash samples, one gave a complete pattern (see Figure 19), and the other two gave partial patterns. Complete patterns were obtained from all three control samples (see Figure 20).

While the foregoing description teaches the principles of the present invention and provides examples for purposes of illustration, those skilled in the art will recognize that various changes in form and details may be made therein without departing from the spirit and scope of the invention.

Claims (25)

1. A method for collecting at least one ridge and valley characteristic marker of an individual, comprising a biological sample containing at least one nucleic acid and its appendages, comprising the following steps: a. Providing at least a first imaging component, said at least the first imaging component being configured to: i. Provide energy waves capable of imaging the at least one ridge and valley feature marker; and ii. Includes a scanning surface configured to allow the energy wave to penetrate the scanning surface; b. A means for providing a substrate assembly having at least a first surface region, wherein the substrate assembly is configured to: i. Allowing the energy waves to penetrate the substrate; ii. Collect the biological sample from the attachment of the individual; and iii. Contains one or more materials configured to be compatible with the reaction conditions of a nucleic acid amplification reaction; c. Position the appendage of the individual on the scanning surface and into contact with the substrate assembly, thereby collecting the biological sample; and d. Collect the at least one ridge and valley feature from the attachment imaged by the energy wave; It also includes the following steps: subjecting the biological sample collected on the substrate assembly to a nucleic acid amplification reaction while it is still present on the substrate assembly. 2. The method of claim 1, wherein the at least first imaging component is an optical scanner or a capacitive scanner. 3. The method of claim 1, wherein the at least one ridge and valley feature marker is collected electronically. 4. The method of claim 1, further comprising the step of: transmitting the at least one ridge and valley feature marker of the individual to at least one database containing the ridge and valley feature marker, the database being selected from the group consisting of: forensic, crime, identity, missing persons, immigration, motor vehicle administration, terrorists, paternity testing, arrested persons, criminal databases, access control, and any combination thereof. 5. The method according to claim 4, wherein the database comprises a child identity database. 6. The method of claim 1, further comprising the step of: providing an identifier on at least one component of the device to associate the biological sample of the individual with at least one ridge and valley feature marker of the individual, wherein the identifier is alphanumeric, graphic, magnetic, or electromagnetic. 7. The method of claim 6, wherein the identifier is a barcode. 8. The method of claim 1, further comprising the step of archiving at least the substrate assembly of the device after performing the step of collecting the biological sample. 9. The method of claim 1, further comprising the step of mailing the device to another location for archiving or testing after the step of collecting the biological sample. 10. The method according to claim 1, wherein the nucleic acid amplification reaction is a polymerase chain reaction. 11. The method of claim 1, further comprising the step of: performing a DNA sequencing reaction while still present on the substrate assembly. 12. The method according to claim 1, wherein the amplification reaction is a component of STR analysis, SNP analysis, or Indel analysis. 13. The method of claim 1, further comprising the step of: placing a mask assembly of the device on the substrate assembly, thereby surrounding the at least first surface region of the substrate assembly, wherein the mask assembly comprises one or more materials compatible with the reaction conditions of the nucleic acid amplification reaction. 14. The method of claim 13, wherein the step of placing the mask assembly on the substrate assembly is performed prior to the step of collecting the biological sample. 15. The method of claim 13, wherein the step of placing the mask assembly on the substrate assembly is performed after the step of collecting the biological sample and before the step of subjecting the biological sample on the substrate assembly to a nucleic acid amplification reaction. 16. The method of claim 1, further comprising the step of sealing the nucleic acid amplification reaction with a sealing solution. 17. The method of claim 1, further comprising the step of: placing the cover assembly of the device on the biological sample collected onto the substrate assembly. 18. The method of claim 17, wherein the step of placing the covering component on the biological sample on the substrate assembly is performed after the reagents for the nucleic acid amplification reaction have been added to the biological sample on the substrate assembly and before the reaction begins. 19. The method of claim 1, wherein the accessory is a finger, toe, palm, or sole. 20. The method of claim 2, wherein the optical scanner comprises an LED, a laser diode, an incandescent light source, or a multispectral imager. 21. The method of claim 1, wherein the at least first imaging component is selected from a camera, an active pixel imager, a CMOS imager, an imager that images in multiple wavelengths, a CCD camera, a photodetector array, and a TFT imager. 22. The method according to any one of claims 1 to 21, wherein the steps of collecting the biological sample and collecting the at least one ridge and valley feature are performed simultaneously. 23. The method according to any one of claims 1 to 18, further comprising the step of: detecting the sequence of the nucleic acid, thereby identifying the individual. 24. The method of claim 23, further comprising the step of: transmitting the nucleic acid sequence identity of the individual to at least one database containing the nucleic acid sequence identity, the database being selected from the group consisting of: forensic, crime, identity, missing persons, immigration, motor vehicle management departments, terrorists, paternity testing, arrested persons, criminal databases, access control, and any combination thereof. 25. The method of claim 24, wherein the database comprises a child identity database.