JP2013113690A - Analysis element, analyzer and analysis method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To accurately detect a sample containing a substance that is the target of analysis.SOLUTION: A reference detection section is provided which detects only the activity of a marked capture substance. Since a signal value corresponding to a maximum capture amount is detected at all the time in the reference detection section, a signal value depending on only factors such as the lapse of time from manufacturing, a storage state, usage environments (temperature, humidity) can be obtained. Therefore, influences of the factors are excluded by comparing the signal value and a signal value of a detecting section.

Description

  The present invention relates to an analysis element, an analysis apparatus, and an analysis method. In particular, the present invention provides an analysis element (chip) for detecting and analyzing a specific component (for example, antigen, antibody, enzyme, substrate, cytokine, etc.) contained in blood, an analysis device, and detecting and analyzing the component. On how to do.

  An immunoassay method using an antigen-antibody reaction is useful as a method used for analysis and measurement in the medical field, biochemical field, or the field of measuring allergens. However, the conventional immunoassay has problems such as a long time for the analysis and complicated operation.

  In recent years, a micro technology (Micro Electro Mechanical System, MEMS) applying a micro processing technology of a semiconductor has been developed. In the biochemical analysis of proteins, genes, etc., micro technology using antigen-antibody reaction (Micro Total Analytical System, μ-TAS) is rapidly developing.

  For example, Patent Document 1 discloses a microchannel-type analyzer having a substrate on a surface of which a microchannel (hereinafter also referred to as “microchannel”) having a width on the order of micrometers is formed. Has been. The analysis apparatus described in Patent Document 1 analyzes a substance to be detected such as an antigen (hereinafter also referred to as “target substance”) using an antibody or an artificial antibody immobilized on a microchannel. It has been proposed to shorten the analysis time or simplify the analysis operation using such an analysis apparatus.

  The structure of the analyzer disclosed in Patent Document 1 is shown in FIG. As shown in FIG. 7, this microchannel type analyzer has a microchannel 111, an introduction hole 112 for introducing a solution into the microchannel 111, and a solution on the surface of a substrate 110 made of a light-transmitting material such as glass or plastic. A liquid storage part 113 for storing the liquid and a discharge hole 114 for discharging the solution from the analyzer are formed. The introduction hole 112 and the discharge hole 114 are respectively provided at both ends of the microchannel 111, and the liquid reservoir 113 is connected to the discharge hole 114. In the microchannel 111, an antibody fixing part 115 is provided. The antibody fixing part 115 has a well-known fixing method (for example, a method using physical adsorption to fix an antibody that specifically binds to a target substance in a solution, an amino group of the antibody, and a functional group of the fixing part. For example, a method of forming a covalent bond between them and fixing them. An antibody is a substance having specific affinity for a target substance.

  FIG. 8 is a diagram for explaining a method of analyzing a target substance using the analyzer shown in FIG. As shown in FIG. 8, the sample solution containing the target substance 120 and the solution containing the labeled antibody 123 are mixed. The labeled antibody 123 is formed by binding an optically detectable labeling substance 121 and an antibody 122 capable of binding to the target substance 120. The labeled antibody 123 and the target substance 120 are combined to form an immune complex 124 (a complex generated by the reaction between the labeled antibody 123 and the target substance 120).

  Next, the solution containing the immune complex 124 is introduced from the introduction hole 112 shown in FIG. When the solution containing the immune complex 124 reaches the antibody fixing part 115, as shown in FIG. 8, the immune complex 124 in the solution and the antibody 125 immobilized on the antibody fixing part 115 are combined. As a result, a complex 126 composed of “immobilized antibody 125—target substance 120—labeled antibody 123” is formed in the antibody fixing unit 115.

  Thereafter, the target substance 120 is detected by optically detecting the labeled substance 121 bound to the labeled antibody 123 in the complex 126 formed by the antibody fixing unit 115. For the detection of the labeling substance 121, for example, according to the type of the labeling substance 121 to be used, a predetermined analytical instrument such as an ultraviolet-visible spectroscopic analysis, a fluorescence analysis, a chemiluminescence analysis, or a thermal lens analysis is used. It is carried out by detecting light absorption, fluorescence or luminescence of the light.

  A calibration curve for the concentration of the target substance 120 is created by applying the target substance analysis method described above to a standard solution containing the target substance 120 with a known concentration. Using this calibration curve, it is possible to measure the concentration of the target substance 120 in a solution whose concentration of the target substance 120 is unknown.

International Publication No. 2006/054689 Pamphlet (released on May 26, 2006)

  When a simple test chip is manufactured using a microchannel chip as disclosed in Patent Document 1, the user can easily store a secondary antibody and a substrate in the chip without setting a reagent in the apparatus. Inspection can be performed. However, due to differences in the storage conditions after chip manufacture, the period from manufacture to use, and / or differences in the environment (temperature, humidity, etc.) used, the measurement results may differ from the actual values. Become. For this reason, there is a risk that the user mistakes a value different from the actual value as a correct test result. Regarding the storage state, the period until use, and the use environment, all conditions are attributed to the user side, and are the conditions that cannot be controlled by the manufacturer of the microchannel chip. Therefore, it has been very difficult for the manufacturer of the microchannel chip to provide the user with a correct value of the test result.

  The present invention has been made in view of the above problems, and its purpose is to make the user aware of the storage state, the period until use, and the usage environment when using the microchannel chip storing the reagent. Without providing a correct value as a detection result.

  In order to solve the above-described problems, an analytical element according to the present invention captures a fluid and a first capturing material that captures a substance to be detected in the fluid and has a labeling substance bound thereto. A channel, a detection unit in which a second capture substance for capturing the substance to be detected in the fluid is disposed, and a detection unit provided in the flow path. And a reference detection unit in which a third capture substance for capturing the first capture substance is disposed, wherein the third capture substance is obtained by saturated adsorption. The first capturing substance is captured. In other words, in the above-described configuration, the third capture substance captures the first capture substance with respect to the flow path so that the maximum amount of the first capture substance that can be adsorbed by the third capture substance is captured. Material is being washed away.

  The signal value of the detection unit varies depending on the concentration of the target substance (substance to be detected) contained in the sample, and the value differs from the actual signal value depending on the storage environment and use environment. However, with the above configuration, since the signal value of the reference detection unit always detects a signal value corresponding to the amount of the first capture substance captured by saturation adsorption (always a constant amount), the reference detection unit It is possible to eliminate the influence of the above factors by referring to the signal value.

  For example, the signal (for example, fluorescence) emitted from the labeling substance bound to the first capture substance varies depending on various conditions. For example, if the labeling substance becomes old, the signal emitted by the labeling substance becomes weaker with time. That is, the signal emitted by a certain amount of labeling substance is not always constant. At this time, with the above configuration, since the third capture substance can always bind a certain amount of the first capture substance, even if the labeling substance is old, it is constant at the time of detection. A signal (eg, fluorescence intensity) per amount of the first capture material can be detected. And based on the said signal (signal per fixed amount of 1st capture | acquisition substances), by correcting the signal detected by the said detection part, of the 1st capture | acquisition substance captured by the said detection part The amount can be calculated accurately. Since the first capture substance captures the substance to be detected in the fluid, if the amount of the first capture substance captured by the detection unit can be accurately calculated, the first capture substance is captured by the detection unit. The amount of substance to be detected can also be accurately calculated.

  In the analysis element according to the present invention, it is preferable that the reference detection unit is provided on the downstream side of the detection unit.

  If it is the said structure, since a 1st capture substance passes through a detection part and is sent to a reference detection part, it will give priority to reaction of a detection part (capturing of the substance which should be detected by a 2nd capture substance) It becomes possible to make it. In addition, since the target substance can also be reacted at the detection unit before passing through the reference detection unit, the influence of non-specific adsorption or the like on the reference detection unit can be reduced.

  In the analytical element according to the present invention, the amount of the first capture substance flowing in the flow path is an amount equal to or greater than the sum of the amount of the second capture substance and the amount of the third capture substance. Is preferred.

  With the above configuration, the amount of the first trapping substance can be easily adjusted because the amount of the first trapping substance is satisfied so that the reference detection unit always captures saturated adsorption. it can. Moreover, according to the said structure, a target substance can be measured more correctly.

  In the analytical element according to the present invention, the flow path is branched, and the detection unit is provided in one of the branched flow paths, and the flow path is different from the branched flow path. It is preferable that the reference detection unit is provided.

  If it is the said structure, since a reference detection part and a detection part can react independently, respectively, it becomes possible to detect without affecting each other.

  In the analysis element according to the present invention, the amount of the first capture substance flowing in the flow path is the amount of the second capture substance in the detection unit or the amount of the third capture substance in the reference detection unit. It is preferable that it is 2 times or more.

  With the above configuration, the amount of the first capture substance can be easily adjusted because the amount of the first capture substance is satisfied so that the reference detection unit always captures the saturated adsorption. . Moreover, according to the said structure, a target substance can be measured more correctly.

  The analytical element according to the present invention preferably includes a first storage unit for storing the first capture substance and introducing the first capture substance into the flow path.

  If it is the said structure, a user can detect, without preparing a 1st capture substance separately. By storing the first capture substance in the chip, the detection signal depends on the passage of time and the storage state after manufacturing the analytical element. Can be ignored.

  The analytical element according to the present invention is preferably provided with a second storage unit for storing a chemical substance that reacts with the labeling substance and for introducing the chemical substance into the flow path.

  If it is the said structure, a user can detect, without preparing the chemical substance which reacts with a labeling substance separately. As described above, the detection signal depends on the passage of time and the storage state after the analysis element is manufactured, but can be easily calibrated by the reference detection unit.

  In the analysis element according to the present invention, it is preferable that an introduction portion for receiving the fluid and introducing the received fluid into the flow path is provided.

  If it is the said structure, since an introducing | transducing part exists in an analysis element, it becomes possible to send liquid easily.

  In the analysis element according to the present invention, it is preferable that the reference detection unit and the detection unit include the same detection means.

  If it is the said structure, in order to detect by the same detection means, even if the detector itself is influenced by the change of use environments, such as heat, the influence can be excluded.

  In the analysis element according to the present invention, it is preferable that the reference detection unit and the detection unit each include a working electrode and a reference electrode.

  If the said structure is used, it will become possible to detect a target substance electrochemically in a detection part. The target substance to be detected electrochemically may be one that is electrochemically active per se, or one that is modified with an electrochemically active substance. As a method for electrochemically detecting a target substance, a current value obtained from an electrochemically active substance may be measured with a detection electrode.

  In the analytical element according to the present invention, it is preferable that the working electrode and the reference electrode have the same size.

  With the above configuration, since the electrodes having the same size are used, it is possible to easily compare the signal values without considering the size of the diffusion layer spreading on the electrode interface.

  In the analysis element according to the present invention, it is preferable that the detection unit and the reference detection unit are formed of a light-transmitting material.

  According to the above configuration, the target substance can be optically detected by the detection unit. The target substance to be detected optically may be one that has optical characteristics or may be modified with a substance that has optical characteristics. Examples of the optical characteristics include light absorption characteristics, light emission characteristics, and color development characteristics. Note that the emission includes fluorescence. Examples of the substance having optical characteristics include a light absorbing dye, a light emitting dye, and a coloring dye. As a method for optically detecting the target substance, any method can be used as long as it detects the above-described optical characteristics. Examples of such methods include conventionally known methods such as ultraviolet-visible spectroscopy, fluorescence analysis, chemiluminescence analysis, and thermal lens analysis. If a target substance having optical characteristics is used, quantitative measurement can be performed by measuring the optical characteristics (measurement of chemiluminescence amount change, fluorescence change, and absorbance change).

  In the analytical element according to the present invention, the first capture substance is preferably an antibody against the substance to be detected.

  A substance to be detected by a microchannel type analyzer used for biochemical analysis is often an in vivo protein. Antibodies that are difficult to denature are optimal substances for use as capture substances.

  In the analytical element according to the present invention, the second capture substance is preferably an antibody against the substance to be detected.

  A substance to be detected by a microchannel type analyzer used for biochemical analysis is often an in vivo protein. Antibodies that are difficult to denature are optimal substances for use as capture substances.

  In the analytical element according to the present invention, the third capture substance is preferably an antibody against the first capture substance.

  A substance to be detected by a microchannel type analyzer used for biochemical analysis is often an in vivo protein. Antibodies that are difficult to denature are optimal substances for use as capture substances.

  In the analytical element according to the present invention, the labeling substance is preferably an enzyme.

  If it is the said structure, since an enzyme is used as a labeling substance, the quantity of the 1st capture | acquisition substance can be easily detected by carrying out an enzyme substrate reaction.

  In the analytical element according to the present invention, the labeling substance is preferably a fluorescent material.

  If it is the said structure, since a fluorescent material is used as a labeling substance, it becomes possible to detect simply by carrying out a direct fluorescence detection.

  In the analytical element according to the present invention, the chemical substance is preferably a substrate that reacts with an enzyme.

  If it is the said structure, it becomes possible to detect a target substance simply by enzyme substrate reaction.

  In the analysis element according to the present invention, a barcode in which information relating to the correlation between the detection signal value derived from the signal value of the detection unit and the signal value of the reference detection unit and the concentration of the substance to be detected is written Alternatively, an IC tag is preferably provided.

  With the above configuration, information can be written in advance on the chip. The information to be written can be changed according to the lot at the time of manufacture and the lot of the reagent.

  In order to solve the above-described problems, an analysis apparatus according to the present invention is characterized by including the analysis element according to the present invention.

  If it is the said structure, since the value corresponding to the 1st capture | acquisition substance capture | acquired so that it may become saturation adsorption | suction is surely detected for the signal value of a reference detection part, by referring the signal value of a reference detection part, It becomes possible to eliminate the influence of the usage environment.

  In the analyzer according to the present invention, it is preferable to calculate the concentration of the substance to be detected from a relative signal value obtained by comparing the signal value of the detection unit and the signal value of the reference detection unit.

  If it is the said structure, only the signal value depending on the density | concentration of a target substance will be obtained by offsetting the influence by the time passage after a chip manufacture, a preservation | save state, and a use environment, for example by taking a ratio with the signal value of a reference detection part. It becomes possible. In addition to the ratio, any correction means such as a difference or a correction coefficient may be used.

  The analyzer according to the present invention preferably issues a warning when the signal value of the reference detection unit is equal to or less than a reference value.

  With the above configuration, it is possible to notify the user that a detection error has occurred by issuing a warning without displaying unreliable data to the user.

  In the analysis apparatus according to the present invention, it is preferable that the detection means for detecting the signal value of the detection unit and the signal value of the reference detection unit are the same.

  With the above configuration, since the same detection means is used, a common detector can be used for the signal value of the detection unit and the signal value of the reference detection unit, thereby simplifying the configuration of the analyzer. be able to.

  The analyzer according to the present invention is preferably provided with a reader for barcode or IC tag.

  If it is the said structure, it will become possible to read information from the barcode provided in the analysis element or the IC tag. The read information can be used for calibration.

  The analysis method according to the present invention calculates the detection signal value by comparing the signal value of the reference detection unit and the signal value of the detection unit using the analysis element of the present invention in order to solve the above problem. It is characterized by that.

  With the above configuration, the concentration of the target substance can be determined by offsetting the influence of the elapsed time, storage state, and usage environment after chip manufacture by taking the ratio of the signal value of the reference detection unit to the signal value of the detection unit. It is possible to obtain only signal values depending on the.

  In the analysis method according to the present invention, the detection signal value is preferably calculated from a ratio between the signal value of the detection unit and the signal value of the reference detection unit.

  With the above configuration, it depends on the concentration of the target substance by offsetting the influence of the elapsed time, storage state, and usage environment after chip manufacture by taking the ratio of the signal value of the reference detector to the signal value of the detector. Only the obtained signal value can be obtained.

  In the analysis method according to the present invention, it is preferable to determine a detection error when the signal value of the reference detection unit is equal to or less than a reference value.

  If it is the said structure, a detection error can be conveyed to a user, without displaying unreliable data to a user.

  In the analysis method according to the invention, a correlation between the detection signal value and the concentration of the substance to be detected is input to the analysis element in advance, and the concentration of the substance to be detected is determined from the detection signal value and the correlation. Is preferably calculated.

  If it is the said structure, since a detection signal value is calculated using a reference detection part and a detection part, it becomes possible to make a value close | similar to an actual value into an output.

  In the analysis method according to the present invention, it is preferable to correct the correlation based on information on at least one of the manufacturing lot and the reagent lot of the analytical element.

  If it is the said structure, it will become possible to correct | amend the said difference of the correlation produced by the difference of a manufacturing lot and a reagent lot.

  In the analysis method according to the present invention, it is preferable that the information is read from a barcode or an IC tag provided on the analysis element.

  If it is the said structure, it will become possible to correct | amend simply, without making a user trouble.

  According to the present invention, the original accurate value can be detected without depending on the storage state after manufacturing the microchannel chip, the period until use or the use environment.

  According to the present invention, an original accurate value can be detected by an element or device having a simple configuration.

It is a top view of the microchannel type | mold analyzer which concerns on Embodiment 1 of this invention. It is a top view of the microchannel type | mold analyzer which concerns on Embodiment 1 of this invention. It is a top view of the microchannel type | mold analyzer which concerns on Embodiment 1 of this invention. It is a top view of the microchannel type | mold analyzer which concerns on Embodiment 2 of this invention. It is a top view of the microchannel type | mold analyzer which concerns on Embodiment 2 of this invention. It is a top view of the microchannel type | mold analyzer which concerns on Embodiment 2 of this invention. It is the schematic which shows the conventional microchannel type | mold analyzer. It is the schematic which shows reaction in the fixing | fixed part of the antibody provided in the conventional microchannel type | mold analyzer.

  DESCRIPTION OF EMBODIMENTS Hereinafter, embodiments of an analyzer according to the present invention will be described with reference to the drawings, but the present invention is not limited to these. In the following, the present invention is described by taking a microchannel type analyzer as an example. However, the analyzer according to the present invention is not limited to the microchannel type. For example, a microcapillary type analyzer is also included in the present invention. Included in the range.

  In this specification, the term “sample” refers to a specimen (test object) introduced into the introduction unit of the analyzer, and may include a target substance (target substance) to be detected. Also good.

  In this specification, the term “capturing substance” refers to a substance that forms a covalent bond or a non-covalent bond with the target substance by specifically interacting with the target substance. The capture substance is specifically a substance having a relationship between a host and a guest with the target substance. Examples of the capture substance include antigens, antibodies, enzymes, substrates, ligands, receptors, DNA, sugars, peptides And synthetic polymers (for example, molecular imprint polymer).

  In this specification, the terms “upstream” and “downstream” are concepts based on the flow of fluid in the microchannel, and the direction in which the liquid flows is “downstream” and the direction opposite to the flow of the liquid is upstream. Call it. Unless otherwise specified, the introduction portion direction in the flow channel is “upstream”.

[Embodiment 1]
A first embodiment of the present invention will be described with reference to FIG. 1, 2 and 3 are plan views of the microchannel type analyzer according to Embodiment 1 of the present invention.

<1. Microchannel analyzer>
The microchannel type analyzer (microchannel chip) according to this embodiment includes a substrate 100 and a lid 101 (not shown) that overlaps the substrate 100, and a concave fine groove (not shown) is formed on the surface of the substrate 100. A microchannel 2) is formed. Further, a fine through hole (microchannel 2) may be formed in the substrate 100 without the lid 101. In the present specification, “fine” is intended to have a diameter on the order of μm, and specifically, a size that can be formed using a semiconductor microfabrication technique. The

  The microchannel 2 defines the flow path of the analyzer according to the present embodiment. On the surface of the substrate 100, an introduction part 1 for receiving a fluid to be introduced and a discharge part 7 for discharging the fluid from the flow path are further formed, and are connected to both ends of the microchannel 2, respectively. That is, the microchannel 2 connects the introduction part 1 and the discharge part 7 on the surface of the substrate 100. The introduction part 1 may be a part that stores fluid introduced into the microchannel 2, and the discharge part 7 may be a part that stores fluid discharged from the microchannel 2. In the present specification, the boundary portion between the microchannel 2 and the introduction portion 1 or the discharge portion 7 is referred to as an introduction hole or a discharge hole (not shown) as necessary.

  By overlaying the lid 101 on such a substrate 100, the microchannel 2 is isolated from the outside of the substrate. However, first and second through holes (not shown) penetrating the substrate 100 or the lid 101 communicate the introduction portion 1 and the discharge portion 7 with the outside of the substrate, respectively. Thereby, the fluid can be supplied to the microchannel 2 from the outside of the substrate, and the fluid can be discharged from the microchannel 2 to the outside of the substrate.

  Further, a detection unit 3 for detecting a substance in the fluid flowing in the microchannel 2 is provided in the microchannel 2. A second capture substance 4 that captures a substance to be detected and analyzed is immobilized on the detection unit 3. Further, in the microchannel 2, a reference detection unit 5 for evaluating the activities of the labeled first capture substance and chemical substance stored in the analyzer is provided. A third capture substance 6 that captures the first capture substance is immobilized on the reference detection unit 5. The detection unit 3 and the reference detection unit 5 may be provided anywhere within the microchannel 2, but preferably, the detection unit 3 is provided on the introduction unit 1 side, and the reference detection unit 5 is provided on the discharge unit 7 side. It is preferable from the viewpoint of reactivity.

  Although not shown, in the analyzer according to the present embodiment, the driving means for promoting the movement of the fluid in the microchannel 2 from the introduction unit 1 to the discharge unit 7 is performed by the introduction unit 1 and the discharge unit 7. It may be connected to at least one. Examples of such driving means include an extrusion pump and a suction pump. When the fluid is fed into the microchannel 2 using the extrusion pump, the extrusion pump may be connected to the introduction unit 1. When the fluid is drawn out from the microchannel 2 using the suction pump, the suction pump is discharged to the discharge unit 7. Can be linked to. In addition to using the pump as described above, the solution can be flowed by a conventionally known method (for example, a method using a capillary phenomenon or a method using a water-absorbing substance).

  The fluid provided to the analyzer according to the present embodiment may be a gas or a liquid, but is preferably a liquid when used for biochemical analysis by a micro technology.

  Hereinafter, members included in the analyzer according to the present embodiment will be described in detail.

<1.1 Substrate>
As the substrate 100 and the lid 101, for example, an insulating substrate can be used. Examples of the substrate having an insulating property include a silicon substrate, a quartz substrate, an aluminum oxide substrate, a glass substrate, and a plastic substrate on which an insulating material such as an oxide film is formed.

  In the case of optically detecting a substance, a light transmissive substrate can be used as the substrate 100 or the lid 101. Examples of the light transmissive substrate include a glass substrate, a quartz substrate, and a substrate made from a light transmissive resin.

  In addition, when a substance is detected using chemiluminescence, a glass or plastic material having low spontaneous fluorescence and transparency (for example, polyimide, polybenzimidazole, polyetheretherketone, polysulfone, polyetherimide, polysiloxane) Ethersulfone or polyphenylene sulfite can also be used as the substrate 100 or the lid 101.

  Note that the thickness of the substrate 100, which is preferable for use in a microchannel analyzer, is about 0.1 to 5 mm. The lid 101 may have the same thickness as the substrate 100 or may be thinner than the substrate 100.

<1.2 Microchannel>
The depth of the microchannel 2 formed on the surface of the substrate 100 is preferably about 0.1 to 1000 μm, and the width is preferably about 0.1 to 1000 μm, but is not limited thereto. The length of the microchannel 2 can be appropriately designed according to the size of the substrate 100, and is preferably about 50 to 800 μm.

  The flow path of the microchannel 2 may have a prismatic shape or a cylindrical shape along the direction in which the fluid flows. That is, the shape of the cross section of the microchannel 2 perpendicular to the direction in which the fluid flows may be a rectangle, a trapezoid, or a circle (semicircle).

  The microchannel 2 can be manufactured by forming irregularities on the substrate 100, for example. For example, a recess may be formed on the substrate 100, and the recess may be the microchannel 2. Alternatively, a plurality of protrusions may be formed on the substrate 100, and a region surrounded by these protrusions may be the microchannel 2. . Alternatively, the microchannel 2 may be formed from a combination of a concave portion and a convex portion, and a combination of the concave portion and the convex portion.

  Examples of the method for forming irregularities on the substrate 100 include a method by machining as a direct processing method, a method by laser processing, an injection molding using a mold, a press molding, and a casting method. Injection molding using a mold is particularly suitable because it is excellent in mass productivity and has high shape reproducibility. In addition, when the material of the substrate 100 is silicon, glass, or the like, the pattern of the microchannel 2 on the substrate 100 can be formed by a photolithography method or an etching method.

  The introduction unit 1 and the discharge unit 7 are illustrated as previously formed on the substrate 100, but are limited as long as the microchannel 2 and the outside of the substrate are connected via the introduction unit 1 and the discharge unit 7. For example, you may form as a 1st and 2nd through-hole. The sizes of the introduction unit 1 and the discharge unit 7 can be appropriately changed according to the size and shape of the microchannel 2. However, in order to use the analyzer according to the present embodiment as a microchannel type analyzer, the diameter is small. It is preferable that it is 10 micrometers or more. The introduction part 1 and the discharge part 7 formed as separate members from the substrate 100 are arranged outside the substrate, and the introduction part 1 and the discharge part 7 are respectively connected to the introduction hole and the discharge hole through the first through hole and the second through hole. A connected mode may be used.

<1.3 Detection unit>
As described above, the detection unit 3 is a part that detects a substance in the fluid flowing through the microchannel 2. As shown in FIG. 1, the detection unit 3 includes a substance to be detected and analyzed (hereinafter referred to as a target for analysis). A second capture substance 4 that captures the target substance is immobilized. The second capture substance 4 is a substance (for example, an antibody, an antigen, a peptide, DNA, a sugar, a synthetic polymer (for example, a molecular imprint polymer), an enzyme, a substrate, a ligand, or a receptor that has a host-guest relationship with the target substance. In particular, an antibody or a synthetic polymer is preferable because its activity is stable. In addition, as a method for immobilizing the second capture substance 4, a known method such as a physical adsorption method, a chemical bonding method, or a covalent bonding method can be appropriately employed.

  The structure of the detection part 3 is not specifically limited, It can determine suitably with the detection method of a target substance. When the target substance is optically detected based on absorbance or light emission (including fluorescence), the light-transmitting portion of the microchannel 2 may be used as the detection unit 3. In order to simplify the manufacture of the analyzer, it is preferable to form the entire substrate 100 or the entire lid 101 with a light transmissive material such as glass, quartz, or a light transmissive resin. In such a case, as shown in the figure, the second capture substance 4 may be immobilized on the inner wall surface of one region of the microchannel 2 that functions as the detection unit 3.

  In addition, when the target substance is detected electrochemically, the detection unit 3 may include a detection unit having a detection electrode formed in the microchannel 2. The detection electrode only needs to be composed of at least two electrodes, that is, a reference electrode and a working electrode, but is preferably composed of three electrodes provided with a counter electrode in addition to the reference electrode and the working electrode. 1 to 3 show a configuration in which the second capture substance 4 is immobilized on the inner wall surface of one region of the microchannel 2 that functions as the detection unit 3, but when a detection electrode is used, It is only necessary that the second capture substance 4 is immobilized on at least the working electrode.

  The reference electrode, the working electrode, and the counter electrode can be formed in the microchannel 2 by a microfabrication technique using a conventional photolithography technique. As the conductive material of the electrode, for example, gold, platinum, silver, chromium, titanium, iridium, copper, or carbon can be used. As the reference electrode, a silver / silver chloride electrode is preferably used from the viewpoint of stability of the standard potential.

<1.4 Reference detection unit>
As described above, the reference detection unit 5 is a site for evaluating the activities of the labeled first capture substance and chemical substance stored in the chip. As shown in FIG. 1, a third capture substance 6 for capturing the first capture substance is immobilized on the reference detection section 5 provided between the detection section 3 and the discharge section 7. .

  Similarly to the second capture substance 4, the third capture substance 6 is a substance having a host-guest relationship with the first capture substance (for example, antigen, antibody, enzyme, substrate, ligand, receptor, DNA, sugar , A peptide or a synthetic polymer (for example, a molecular imprint polymer), and the like. In particular, an antibody or a synthetic polymer is preferable because its activity is stable. As a method for immobilizing the third capture substance 6, a known method such as a physical adsorption method, a chemical bonding method, or a covalent bonding method can be appropriately employed.

  The configuration of the reference detection unit 5 is the same as that of the detection unit 3, and is not particularly limited. The reference detection unit 5 can be appropriately determined depending on the detection method of the first capture substance. When the first capture substance is optically detected based on absorbance or light emission (including fluorescence), the light transmissive portion of the microchannel 2 may be used as the reference detection unit 5. In order to simplify the manufacture of the analyzer, the entire substrate 100 or the entire lid 101 is preferably made of a light-transmitting material such as glass, quartz, and a light-transmitting resin. In such a case, as shown in the drawing, the third capture substance 6 may be immobilized on the inner wall surface of one region of the microchannel 2 that functions as the reference detection unit 5. In addition, when the first capture substance is detected electrochemically, the reference detection unit 5 only needs to include detection means having a detection electrode formed in the microchannel 2. The detection electrode only needs to be composed of at least two electrodes, that is, a reference electrode and a working electrode, but is preferably composed of three electrodes provided with a counter electrode in addition to the reference electrode and the working electrode.

  In addition, the configuration of the reference detection unit 5 is preferably the same detection means and / or detection structure as the detection unit 3. If the detection means and / or the detection structure are the same, the influence of detection variation can be ignored. Further, since the detection signal value also varies depending on the temperature, if the detection means and / or the detection structure are the same, the fluctuation range can be regarded as the same.

<1.5 Labeled first capture substance>
The labeling substance of the labeled first capture substance for detecting the substance in the fluid may be any substance that can be used as a labeling agent, such as a dye, a fluorescent dye (or fluorescent protein), an enzyme, or a radioactive substance. As a capture substance, a substance having a host-guest relationship as described above (for example, antigen, antibody, enzyme, substrate, ligand, receptor, DNA, sugar, peptide, or synthetic polymer (for example, molecular imprint polymer) In particular, an antibody or a synthetic polymer is preferable because its activity is stable.
<1.5.1 Storage Method of Labeled First Capture Substance>
The storage method of the labeled first capture substance in the chip may be any of storage in liquid and dry storage, and can be selected as appropriate according to the shape of the chip and the characteristics of the reagent. Further, the storage location (first storage unit) may be any portion between the introduction unit 1 and the detection unit 3. For example, a storage unit (first storage unit) may be provided in the flow path, in the introduction unit 1 or on the detection unit 3. The first capture substance is preferably stored after the treatment with a blocking agent on the wall surface in the chip or during the treatment. An existing method can be used as the blocking method. For example, the flow path substrate itself may be made of a material having a high blocking ability, or the blocking treatment can be performed by immersing in a blocking solution such as BSA.

<1.5.2 Channel structure>
As the flow channel structure, a branched flow channel may be prepared in the microchannel as shown in FIG. 2, and the labeled first capture substance may be stored in the branched first capture substance flow path 10. With this structure, the sample liquid and the first capture substance can be fed to the detection unit 3 and the reference detection unit 5 without contamination. Further, it is preferable to provide a valve 9 in the first capture substance flow path 10. If it is the said structure, it will become possible to prevent the contamination at the time of sample feeding better. However, the position of the valve 9 shown in FIG. 2 is not particularly limited. It is only necessary to exhibit a function of preventing the liquid from flowing into the branched first capture substance flow channel 10 when the sample is fed from the introduction unit 1. For example, a structure for physically stopping the flow of fluid in the first capture substance flow path 10, a structure for cutting the fluid in the first capture substance flow path 10, and the first capture substance flow path 10 It may have a structure for separating the fluid in the inside, and may have all functions as necessary. Examples of the preferable valve 9 include a rotary screw type valve, a detachable weir plate, closing by pressure, or cutting of liquid by gas control.

<1.5.3 Amount of labeled first capture substance>
The storage amount of the labeled first capture substance is an amount that allows the first capture substance to react sufficiently in the detection unit 3 and an amount that can be reacted in a substantially saturated manner in the reference detection unit 5.

  Specifically, when the immobilized amount of the second capture substance 4 in the detection unit 3 is obtained as data, the amount of the first capture substance can be expressed by the following equation.

(Storage amount of the first capture substance) ≧ (immobilization amount of the second capture substance 4) + (immobilization amount of the third capture substance 6)
However, when a capture substance that captures a large number of epitopes with respect to one target substance such as a polyclonal antibody is used as the first capture substance, it is desirable to store the first capture substance labeled about three times as much as the above formula. .

(Stored amount of the first capture substance) ≧ {(immobilization amount of the second capture substance 4) + (immobilization amount of the third capture substance 6)} × 3
When the immobilized amount data of the second capture substance in the detection unit 3 is not obtained, the first capture substance can be prepared as follows. Specific preparation examples are shown below.
(1) Confirmation of Reference Detection Unit 5 Using the first capture substance (standard substance) whose concentration is known, the detection range of the first capture substance in the reference detection unit 5 is examined. By deriving the product of the obtained saturated concentration and the delivered volume, the reference detector 5 can calculate the amount necessary for the first trapping substance to be saturated and adsorbed.
(2) Confirmation of the detection unit 3 Using the target substance (standard material) whose concentration is known, the maximum amount of the first capture substance necessary in the detection unit 3 is examined. Specifically, in the detection unit 3 that is reacted at the assumed maximum concentration of the target substance, the first trapping substance concentration necessary for saturated adsorption with respect to the target substance trapped on the detection unit 3 is examined. By deriving the product of the obtained saturated concentration and capacity, the amount necessary for the first trapping substance to be saturated and adsorbed can be calculated.

  As mentioned above, more than the sum of the quantity of the 1st capture substance obtained from (1) and (2) is preserve | saved in the preservation | save part (1st preservation | save part) in a microchannel.

<1.6 Analysis method>
An example of an analysis method using the analysis chip according to this embodiment is shown below. The driving means for flowing the fluid in the microchannel 2 includes a method using an extrusion pump connected to the introduction unit 1, a method using a suction pump connected to the discharge unit 7, and a method using capillary force and / or a water-absorbing substance. Either is acceptable.

(1) Sample Introduction and Reaction As a first example, in the microchannel chip shown in FIG. 1, a storage unit (first storage unit) in which a labeled first capture substance is stored is used as an introduction unit. An analysis example regarding the chip provided in 1 will be described. A sample is introduced from the introduction unit 1 into the microchannel 2. The target substance in the sample first flows into the detection unit 3 while reacting with the first capture substance stored in the introduction unit 1. Since the target substance in the flowed sample also reacts with the second capture substance 4 on the detection unit 3, as a result, the second capture substance 4 on the detection unit 3 captures the target substance bound to the first capture substance. Will be. On the other hand, the excessive first capture substance that has flowed to the reference detection unit 5 is captured by the third capture substance 6 on the reference detection unit 5.

  As a second example, in the microchannel chip illustrated in FIG. 2, a storage unit (first storage unit) in which the first capture material is stored is provided in the first capture material channel 10. The chip will be described. A sample is introduced from the introduction unit 1 into the microchannel 2. The target substance that has flowed into the detection unit 3 is captured by the second capture material 4 on the detection unit 3. At this time, it is preferable that the valve 9 be closed so that the liquid does not flow into the first capture substance flow path 10. Next, the valve 9 is opened, the buffer is sent from the introduction part 11, and the first capture substance stored in the first capture substance flow path 10 is sent to the microchannel 2. The sent first capture substance reacts with the target substance captured on the detection unit 3 and the third capture substance 6 on the reference detection unit 5, and is captured by each. In addition, a washing operation for washing the microchannel 2 may be performed after feeding the sample and before feeding the first capture substance. In the cleaning operation for cleaning the microchannel 2, the cleaning liquid may be sent from the introduction unit 1 to the microchannel 2, or from a newly provided branch channel (not shown) of the microchannel 2. You may send a washing | cleaning liquid.

(2) Detection of target substance When the labeling substance of the first capture substance is a labeling substance that can be measured directly, such as a fluorescent dye, the labeling substance can be detected after the first capture substance is introduced. It is. Alternatively, the labeling substance may be detected after the cleaning liquid is fed from the introduction unit 1. Similarly to the above, the cleaning liquid may be sent from a dedicated cleaning channel (not shown).

  When the labeling substance of the first capturing substance is an enzyme or the like, it is a labeling substance that requires a reaction with a chemical substance (corresponding to a substrate in the case of an enzyme) for detection. In that case, for example, in the microchannel chip shown in FIG. 3, detection is performed by sending a solution containing a chemical substance from the chemical substance flow path 13 provided with the second storage unit in which the chemical substance is stored. It is possible to carry out a reaction for Moreover, you may put in washing | cleaning operation before sending the solution containing a chemical substance. For example, when the labeling substance of the first capture substance is an ALP enzyme and pAPP is used as a substrate, pAPP changes to pAP by the reaction between the enzyme and the substrate. Note that pAP is an electrochemically active substance. Since the substrate changes to an electrochemically active substance, a signal value can be acquired as a current value.

The detection is performed by the detection unit 3 and the reference detection unit 5. The detection unit 3 detects the amount of the first captured substance captured on the detection unit 3 (corresponding to the amount of the captured object), and the reference detection unit 5 detects the first captured on the reference detection unit 5. Detect the amount of trapped material.
<1.7 Correction of signal value>
In the case of a conventional microchannel chip, calibration is performed from the signal value of the detection unit 3. The detection signal value is expressed as follows.

・ Detected signal ∝ Immobilization amount of capture substance (second capture substance) x Reaction rate of target substance x Reaction rate of labeled capture substance (first capture substance) x Enzyme substrate reaction rate (Use enzyme substrate reaction) X detection efficiency Immobilization amount, reaction rate, and detection efficiency are as follows: (A) Production lot (capture substance, labeling substance, etc.), (B) Immobilization amount or storage amount, (C) Use environment (humidity, The signal value may change depending on the temperature and the time after manufacture, which may cause a difference from the actual value.

  The example of the factor which changes a signal value concretely is described below.

  Since (A) and (B) above depend on manufacturing conditions, it can be dealt with by correcting the device for detecting the microchannel chip in advance. However, (C) the usage environment is a condition that depends on the user, so the manufacturer of the microchannel chip cannot control it.

  In addition to the above factors, the signal value of the detection unit 3 varies depending on the concentration of the target substance contained in the sample, but the signal value of the reference detection unit 5 is always detected by a signal value corresponding to saturated adsorption. Therefore, only the influence of the above factors can be considered. Specifically, the signal value of the reference detection unit 5 can be expressed as follows.

・ Signal value of the reference detector ∝ Amount of immobilized capture substance (third capture substance) × reaction rate of labeled capture substance (first capture substance) × enzyme substrate reaction rate (when using enzyme substrate reaction) ) × detection efficiency Compared with the signal value equation of the detection unit 3 described above, it is possible to cancel out the factors affected by the use environment, so by comparing the signal value of the detection unit 3 with the signal value of the reference detection unit 5 It is possible to reduce the difference from the actual value. The correction method can be performed by taking a ratio, but any other correction means such as applying a difference or a correction coefficient may be used.

  For example, when the enzyme activity is reduced by 20% due to the storage time, the signal value of the detection unit 3 may be reduced by a maximum of 20%, but the signal value of the reference detection unit 5 is also reduced. When compared, the effect of enzyme activity is negligible.

  Regarding the corrections (A) and (B), there is a possibility that the signal value changes depending on the lot of the manufacturing chip. Therefore, when the lot is changed, the user can respond by inputting correction information in advance to an apparatus or software for detecting a microchannel chip. In addition, it is possible to embed information in the chip by adding an IC tag, a barcode, or the like to the chip, read the information on the detection device side, and automatically correct the information. For example, it is possible to perform calibration from the detection signal value obtained as a detection result by inputting a calibration curve, data information, etc. in advance on the device side. Since there is a possibility that the calibration curve differs depending on the difference, information for calibration can be input manually or in a bar code or IC tag. The apparatus can correct the calibration curve or data based on the input information. Any known technique can be used as a method of reading a barcode or IC tag by the apparatus.

<1.8 Detection error>
The reference detection unit 5 can also be used to determine whether or not the detection by the microchannel chip has failed. The reference detection unit 5 detects a signal value when the labeled first capture substance is saturated and adsorbed, but when the signal value is detected below a predetermined value, the reliability of the measurement itself is low. It is possible to judge. The predetermined value can be used as a reference value. When the signal value of the reference detection unit 5 falls below the standard value at the time of detection, a warning is issued to the user and the calibration value is not displayed to prevent the user from telling the erroneous calibration value. Is possible. The warning may be a light warning or a sound warning, and a conventionally known method can be used. However, when detection by light is performed in the detection unit and the reference detection unit, a warning by sound is preferable so as not to affect them.

[Embodiment 2]
A second embodiment of the present invention will be described with reference to FIGS. In addition, about the same structure as Embodiment 1, the detailed description is abbreviate | omitted here.

  In the first embodiment, the reference detection unit 5 and the detection unit 3 are arranged in series in the same microchannel, but in the second embodiment, as shown in FIG. 4, the reference detection unit 5 ′ and the detection unit 3 are detected. The unit 3 ′ is arranged in parallel in the branched microchannel. By arranging in parallel, the item of “amount of labeled second capture substance to be stored” is different from that in the first embodiment, but other items are the same as those in the first embodiment. For example, as described in FIGS. 5 and 6, a first capture substance flow path 10 ′, a substrate flow path 13 ′, a cleaning flow path (not shown), and the like can be provided. These can use the same structure as in the first embodiment.

  As shown in FIGS. 4 to 6, the microchannel 2 branches into a plurality of flow paths on the downstream side of the introduction hole 1 ′. The position of the branch point is not particularly limited, but when at least one of the first capture substance flow path 10 ′ and the substrate flow path 13 ′ is provided, these flow paths are connected to the microchannel 2. It is preferable that a branch point exists on the downstream side of the portion to be formed.

  The number of flow paths through which one microchannel 2 branches is not particularly limited and may be set as appropriate. For example, the number may be two, three, four, or more. When the number of branched flow paths is more than two, for example, the number of flow paths can be an even number (for example, 4, 6, 8,...). When the number of branched flow paths is more than two, for example, a plurality of flow paths provided with the detection unit 3 ′ and a plurality of flow paths provided with the reference detection unit 5 ′ may be provided. Is possible. In this case, it is preferable to provide the same number of detection units 3 ′ and reference detection units 5 ′ so that each detection unit 3 ′ and each reference detection unit 5 ′ make a pair. According to the above configuration, it is possible to simultaneously detect and analyze a plurality of types of substances with one microchannel chip. In this case, each detection unit 3 ′ and reference detection unit 5 ′ can be configured according to the substance to be detected and analyzed. That is, the plurality of detection units 3 ′ may be detection units 3 ′ having the same configuration, but may be detection units 3 ′ having different configurations. Similarly, the plurality of reference detection units 5 ′ may be reference detection units 5 ′ having the same configuration, but may be reference detection units 5 ′ having different configurations.

  The size and shape of the cross section of each branched flow path in the direction perpendicular to the direction in which the fluid moves is not particularly limited, but is preferably the same size and shape. According to the above configuration, it is possible to detect and analyze a substance with high accuracy. The size and shape of each branched flow path can be the same as that of the microchannel 2 described in the first embodiment, for example.

  In addition, a valve (not shown) may be provided between the branch point of the flow path and the reference detection unit 5 '. When a sample is introduced into the introduction hole 1 ′ shown in FIG. 4, the sample flows into both branched flow paths. When the valve is provided and the sample is introduced with the valve closed, the target substance flows to the detection unit 3 ′ without flowing to the reference detection unit 5 ′, and is captured by the second capture substance 4 ′. Thereafter, if the first capture substance is introduced into the flow path with the valve opened, the target substance captured on the detection unit 3 ′ and the third capture substance 6 ′ on the reference detection unit 5 ′ Each reacts and the first capture material is captured. If it is the said structure, a target substance can be made to react reliably by the detection part 3 '.

<2.1 Amount of labeled first capture substance>
As in the first embodiment, the storage amount of the labeled first capture substance is an amount that allows the first capture substance to react sufficiently in the detection unit 3 ′, and is almost saturated in the reference detection unit 5 ′. The amount can be reacted.

  Specifically, when the immobilized amount of the second capture substance 4 ′ in the detection unit 3 ′ is obtained as data, the amount of the first capture substance can be expressed by the following formula.

(Immobilization amount of second capture substance 4 ′) ≧ (immobilization amount of third capture substance 6 ′) (preservation quantity of first capture substance) ≧ (immobilization of second capture substance 4 ′) Amount) x 2
(Immobilization amount of third capture substance 6 ′) ≧ (immobilization amount of second capture substance 4 ′) (preservation amount of first capture substance) ≧ (fixation of third capture substance 6 ′) Amount) x 2
Since FIG. 4 shows a two-branch channel, the entire chip is stored twice. In case of n branches, n times are used.

  However, when a capture substance that captures a large number of epitopes for one target substance, such as a polyclonal antibody, is used as the first capture substance, it is desirable to store the first capture substance about three times the above formula.

When the immobilized amount data of the second capture substance 4 ′ in the detection unit 3 ′ is not obtained, the first capture substance can be prepared as follows. Specific preparation examples are shown below.
(1) Confirmation of reference detection unit 5 ′ Using the first capture substance (standard substance) having a known concentration, the detection range of the first capture substance in the reference detection unit 5 ′ is examined. By deriving the product of the capacity from the obtained saturated concentration, the amount necessary for the first capture substance to be saturated and adsorbed in the reference detection unit 5 ′ can be calculated.
(2) Confirmation of detection part 3 'The maximum amount of the first capture substance required in the detection part 3' is examined using a target substance (standard substance) whose concentration is known. Specifically, in the detection unit 3 ′ reacted at the maximum concentration of the target substance assumed, the concentration of the first capture substance necessary for saturated adsorption to the target substance captured on the detection unit 3 ′ is examined. . By deriving the product of the obtained saturated concentration and capacity, the amount necessary for the first trapping substance to be saturated and adsorbed can be calculated.

  As described above, the amount of the first capture substance obtained from (1) and (2) is compared, and the amount twice as large as the larger amount is stored.

  The present invention can also be configured as follows.

  The analysis element of the present invention is provided with a detection unit for detecting a substance in the fluid in the flow path, and the detection unit is provided with a second capture substance for capturing the substance to be detected. In the analysis element using the first capture substance to which the labeling substance is bound, a reference detection unit for detecting the first capture substance is provided in the flow path. A third capture substance that captures one capture substance is disposed, and a maximum amount of the first capture substance is captured with respect to the third capture substance on the reference detection unit.

  In the analysis element of the present invention, it is preferable that the reference detection unit is on the downstream side of the detection unit.

  In the analytical element of the present invention, the necessary amount of the first capture substance is preferably equal to or greater than the sum of the amount of the second capture substance in the detection unit and the amount of the third capture substance in the reference detection unit.

  In the analytical element of the present invention, the flow path is branched on the downstream side of the injection part, the detection part is provided in one branched flow path, and the other branched flow path is It is preferable that the reference detection unit is provided.

  In the analytical element of the present invention, the required amount of the first capture substance is preferably at least twice the amount of the second capture substance in the detection unit or the third capture substance in the reference detection unit.

  In the analysis element of the present invention, the first capture substance is preferably stored in the chip.

  In the analytical element of the present invention, the chemical substance that reacts with the labeling substance is preferably stored in the chip.

  In the analysis element of the present invention, it is preferable to include an injection part that receives a fluid to be injected in the flow path.

  In the analysis element of the present invention, the reference detection unit and the detection unit are preferably detected by the same detection means.

  In the analysis element of the present invention, it is preferable that the detection unit and the reference detection unit each include a working electrode and a reference electrode.

  In the analytical element of the present invention, it is preferable that the working electrode and the reference electrode have the same size.

  In the analysis element of the present invention, it is preferable that the detection unit and the reference detection unit are made of a permeable material.

  In the analytical element of the present invention, the first capture substance is preferably an antibody against the substance to be detected.

  In the analytical element of the present invention, the second capture substance is preferably an antibody against the substance to be detected.

  In the analytical element of the present invention, the third capture substance is preferably an antibody against a labeled capture substance.

  In the analytical element of the present invention, it is preferable that the labeling substance bonded to the first capture substance is an enzyme.

  In the analytical element of the present invention, the labeling substance bonded to the first capture substance is preferably a fluorescent material.

  In the analytical element of the present invention, the standard chemical substance is preferably a substrate that reacts with an enzyme.

  In the analysis element of the present invention, a barcode or IC tag in which information relating to the correlation between the detection signal value derived from the signal value of the detection unit and the signal value of the reference detection unit and the concentration of the substance to be detected is written is provided. Is preferred.

  The analyzer of the present invention is characterized in that the concentration of a substance to be detected is calculated from a relative signal value obtained by comparing the signal value of the detection unit and the signal value of the reference detection unit, using the analysis element of the present invention.

  The analysis device of the present invention is characterized by issuing a warning when the signal value of the reference detection unit is equal to or less than a reference value using the analysis element of the present invention.

  The analysis apparatus of the present invention is an analysis apparatus for detecting the analysis element of the present invention, and is characterized in that the detection means for detecting the detection unit and the reference detection unit are the same.

  The analysis apparatus of the present invention is characterized in that a reader for barcode or IC tag is provided using the analysis element of the present invention.

  The analysis method of the present invention is characterized in that when the detection signal value is calculated using the analysis element of the present invention, the signal value of the reference detection unit and the signal value of the detection unit are compared.

  In the analysis method of the present invention, the detection signal value is preferably calculated from the ratio between the signal value of the detection unit and the signal value of the reference detection unit.

  In the analysis method of the present invention, it is preferable to determine a detection error when the signal value of the reference detection unit is equal to or less than the reference value.

  In the analysis method of the present invention, the correlation between the detection signal value calculated from the signal value of the detection unit and the signal value of the reference detection unit and the concentration of the substance to be detected is input in advance, and the detection obtained from the detection chip It is preferable to output the concentration of the substance to be detected from the signal value.

  In the analysis method of the present invention, it is preferable to correct the correlation by inputting chip information before measurement.

  In the analysis method of the present invention, it is preferable to input the chip information by reading it from a barcode or IC tag provided on the chip and correct the correlation.

  The present invention is not limited to the configurations described above, and various modifications can be made within the scope of the claims, and the technical means disclosed in different embodiments are appropriately combined. The obtained embodiment is also included in the technical scope of the present invention.

  By using the analyzer of the present invention, it becomes possible to display a highly accurate measurement result to the user regardless of differences in use environment and the like. For this reason, the present invention provides a micro-technology using μ-TAS technology, such as chemical microdevices (for example, microchannel chips and microreactors) used for detection of minute chemical substances, and biosensors (for example, allergen sensors). It can be applied to the field using channel chips.

1, 1 'introduction hole 2, 2' microchannel 3, 3 'detection section 4, 4' second capture substance 5, 5 'reference detection section 6, 6' third capture substance 7, 7 'discharge section 9 , 9 'valve 10, 10' first capture substance flow path 11, 11 'introduction hole 12, 12' valve 13, 13 'substrate flow path 14, 14' introduction hole 100, 100 'substrate 101 lid 110 substrate 111 Microchannel 112 Introduction part 113 Liquid reservoir part 114 Discharge part 115 Antibody immobilization part 120 Target substance 121 Label substance 122 Antibody 123 Labeled antibody 124 Immune complex 125 Immobilized antibody 126 complex

Claims (30)

A flow path for flowing a fluid, and a first capture substance that captures the substance to be detected in the fluid and has a label substance bound thereto;
A detection unit that is provided in the flow path and in which a second capture substance for capturing the substance to be detected in the fluid is disposed;
A reference detection unit provided in the flow path and provided with a third capture substance for capturing the first capture substance, and an analysis element comprising:
The analytical element, wherein the third capture substance captures the first capture substance by saturated adsorption.   The analysis element according to claim 1, wherein the reference detection unit is provided on a downstream side of the detection unit.   3. The amount of the first capture substance that flows in the flow path is an amount that is equal to or greater than the sum of the amount of the second capture substance and the amount of the third capture substance. The analytical element described. The flow path is branched,
The detection unit is provided in one of the branched flow paths,
The analysis element according to claim 1, wherein the reference detection unit is provided in a flow channel different from the branched flow channel.   The amount of the first capture substance flowing in the flow path is at least twice the amount of the second capture substance in the detection unit or the amount of the third capture substance in the reference detection unit. The analytical element according to claim 4, wherein the analytical element is characterized in that   The first storage part for storing the first capture substance and introducing the first capture substance into the flow path is provided. 2. The analytical element according to item 1.   The chemical substance that reacts with the labeling substance is stored, and a second storage unit is provided for introducing the chemical substance into the flow path. The analytical element according to item.   The analytical element according to any one of claims 1 to 7, further comprising an introduction portion for receiving the fluid and introducing the received fluid into the flow path.   The analytical element according to claim 1, wherein the reference detection unit and the detection unit include the same detection means.   The analytical element according to claim 1, wherein each of the reference detection unit and the detection unit includes a working electrode and a reference electrode.   The analysis element according to claim 10, wherein the working electrode and the reference electrode have the same size.   The analysis element according to claim 1, wherein the detection unit and the reference detection unit are made of a light-transmitting material.   The analytical element according to claim 1, wherein the first capture substance is an antibody against the substance to be detected.   The analytical element according to claim 1, wherein the second capture substance is an antibody against the substance to be detected.   The analytical element according to claim 1, wherein the third capture substance is an antibody against the first capture substance.   The analytical element according to claim 1, wherein the labeling substance is an enzyme.   The analytical element according to claim 1, wherein the labeling substance is a fluorescent material.   The analytical element according to claim 7, wherein the chemical substance is a substrate that reacts with an enzyme.   A barcode or IC tag is provided in which information relating to the correlation between the detection signal value derived from the signal value of the detection unit and the signal value of the reference detection unit and the concentration of the substance to be detected is written. The analytical element according to any one of claims 1 to 18, wherein the analytical element is characterized in that   An analysis device comprising the analysis element according to claim 1.   21. The analyzer according to claim 20, wherein the concentration of the substance to be detected is calculated from a relative signal value obtained by comparing the signal value of the detection unit and the signal value of the reference detection unit.   The analyzer according to claim 20 or 21, wherein a warning is issued when a signal value of the reference detection unit is equal to or less than a reference value.   23. The analyzer according to claim 20, wherein the detection means for detecting the signal value of the detection unit and the signal value of the reference detection unit are the same.   The analyzer according to any one of claims 20 to 23, wherein a reader for a barcode or an IC tag is provided. Using the analytical element according to any one of claims 1 to 19,
An analysis method comprising: calculating a detection signal value by comparing a signal value of the reference detection unit with a signal value of the detection unit.   26. The analysis method according to claim 25, wherein the detection signal value is calculated from a ratio between a signal value of the detection unit and a signal value of the reference detection unit.   27. The analysis method according to claim 25 or 26, wherein a detection error is determined when a signal value of the reference detection unit is equal to or less than a reference value. The correlation between the detection signal value and the concentration of the substance to be detected is input in advance to the analysis element,
28. The analysis method according to claim 25, wherein the concentration of the substance to be detected is calculated from the detection signal value and the correlation.   29. The analysis method according to claim 28, wherein the correlation is corrected based on information relating to at least one of a manufacturing lot and a reagent lot of the analytical element.   30. The analysis method according to claim 29, wherein the information is read from a barcode or an IC tag provided on the analysis element.