CN111073810A - Microfluidic chip, system and method integrating nucleic acid extraction, amplification and detection - Google Patents
Microfluidic chip, system and method integrating nucleic acid extraction, amplification and detection Download PDFInfo
- Publication number
- CN111073810A CN111073810A CN201911328740.7A CN201911328740A CN111073810A CN 111073810 A CN111073810 A CN 111073810A CN 201911328740 A CN201911328740 A CN 201911328740A CN 111073810 A CN111073810 A CN 111073810A
- Authority
- CN
- China
- Prior art keywords
- nucleic acid
- chamber
- washing
- waste liquid
- acid extraction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 145
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 145
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 145
- 230000003321 amplification Effects 0.000 title claims abstract description 116
- 238000003199 nucleic acid amplification method Methods 0.000 title claims abstract description 116
- 238000000605 extraction Methods 0.000 title claims abstract description 95
- 238000001514 detection method Methods 0.000 title claims abstract description 71
- 238000000034 method Methods 0.000 title claims abstract description 17
- 239000007788 liquid Substances 0.000 claims abstract description 156
- 238000005406 washing Methods 0.000 claims abstract description 133
- 239000002699 waste material Substances 0.000 claims abstract description 97
- 238000003860 storage Methods 0.000 claims abstract description 68
- 239000003480 eluent Substances 0.000 claims abstract description 50
- 238000010828 elution Methods 0.000 claims abstract description 20
- 239000000243 solution Substances 0.000 claims description 22
- -1 polyethylene Polymers 0.000 claims description 11
- 239000004743 Polypropylene Substances 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 10
- 229920001155 polypropylene Polymers 0.000 claims description 10
- 230000008569 process Effects 0.000 claims description 9
- 239000004698 Polyethylene Substances 0.000 claims description 7
- 239000000463 material Substances 0.000 claims description 7
- 229920000573 polyethylene Polymers 0.000 claims description 7
- 239000004793 Polystyrene Substances 0.000 claims description 5
- 239000004676 acrylonitrile butadiene styrene Substances 0.000 claims description 5
- 229920000122 acrylonitrile butadiene styrene Polymers 0.000 claims description 5
- 238000002347 injection Methods 0.000 claims description 5
- 239000007924 injection Substances 0.000 claims description 5
- 239000004417 polycarbonate Substances 0.000 claims description 5
- 229920000515 polycarbonate Polymers 0.000 claims description 5
- 239000004800 polyvinyl chloride Substances 0.000 claims description 5
- 239000000376 reactant Substances 0.000 claims description 5
- 229920002223 polystyrene Polymers 0.000 claims description 3
- 229920000915 polyvinyl chloride Polymers 0.000 claims description 3
- 238000003475 lamination Methods 0.000 claims description 2
- 238000005516 engineering process Methods 0.000 description 18
- 238000011901 isothermal amplification Methods 0.000 description 10
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 238000001917 fluorescence detection Methods 0.000 description 7
- 238000003745 diagnosis Methods 0.000 description 6
- 238000007789 sealing Methods 0.000 description 6
- 238000003825 pressing Methods 0.000 description 5
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 4
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 229920003023 plastic Polymers 0.000 description 4
- 239000004033 plastic Substances 0.000 description 4
- 230000004907 flux Effects 0.000 description 3
- 238000001746 injection moulding Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000001179 sorption measurement Methods 0.000 description 3
- 238000006467 substitution reaction Methods 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 208000035473 Communicable disease Diseases 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000010408 film Substances 0.000 description 2
- 238000002032 lab-on-a-chip Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 208000019331 Foodborne disease Diseases 0.000 description 1
- 208000026350 Inborn Genetic disease Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000034817 Waterborne disease Diseases 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 208000016361 genetic disease Diseases 0.000 description 1
- 238000005459 micromachining Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 238000010137 moulding (plastic) Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 210000000438 stratum basale Anatomy 0.000 description 1
- 238000010104 thermoplastic forming Methods 0.000 description 1
- 238000009757 thermoplastic moulding Methods 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Immunology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Genetics & Genomics (AREA)
- Dispersion Chemistry (AREA)
- Hematology (AREA)
- Clinical Laboratory Science (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
一种集成核酸提取、扩增和检测的微流控芯片、系统及方法，芯片包括进样口、洗涤液存储腔、洗脱液存储腔、核酸提取腔、洗涤废液接收部及恒温扩增腔，核酸提取腔设置有核酸提取滤纸，洗涤液存储腔和洗脱液存储腔分别经单向阀与核酸提取腔相连，核酸提取腔连接洗涤废液接收部和恒温扩增腔，洗涤液存储腔内预存储洗涤液，洗脱液存储腔内预存储洗脱液，洗涤液存储腔用于在外力驱动下使洗涤液突破单向阀流经核酸提取腔进行洗涤，废液流入洗涤废液接收部，洗脱液存储腔用于在外力驱动下使洗脱液突破单向阀进入核酸提取腔进行洗脱，洗脱下的核酸溶液流入恒温扩增腔，废液流入洗脱废液腔。该芯片能够快速、简单、稳定、可靠地完成核酸提取、扩增和检测。
A microfluidic chip, system and method integrating nucleic acid extraction, amplification and detection, the chip includes a sample inlet, a washing liquid storage cavity, an eluent storage cavity, a nucleic acid extraction cavity, a washing waste liquid receiving part and a constant temperature amplification The nucleic acid extraction chamber is provided with nucleic acid extraction filter paper. The washing liquid storage cavity and the eluate storage cavity are respectively connected to the nucleic acid extraction cavity through a one-way valve. The nucleic acid extraction cavity is connected to the washing waste liquid receiving part and the constant temperature amplification cavity. The washing liquid storage The washing liquid is pre-stored in the cavity, and the washing liquid is pre-stored in the washing liquid storage cavity. The washing liquid storage cavity is used to make the washing liquid break through the one-way valve and flow through the nucleic acid extraction cavity for washing under the driving of external force, and the waste liquid flows into the washing waste liquid The receiving part, the eluent storage chamber is used to make the eluent break through the one-way valve and enter the nucleic acid extraction chamber for elution under the driving of external force, the eluted nucleic acid solution flows into the constant temperature amplification chamber, and the waste liquid flows into the elution waste liquid chamber . The chip can quickly, simply, stably and reliably complete nucleic acid extraction, amplification and detection.
Description
Technical Field
The invention relates to the technical field of molecular diagnosis, in particular to a micro-fluidic chip, a system and a method for integrating nucleic acid extraction, amplification and detection.
Background
Micro-fluidic chips, also known as Lab-on-a-Chip (LOC), have rapidly grown up after Micro-Electro-mechanical systems (MEMS) technology was introduced in the 90 s of the 20 th century, and received much attention from the scientific community. The microfluidic chip technology is an experimental technology for performing precise control on microfluid in a microchannel, has the advantages of low consumption, low cost, high flux, automatic operation and the like, is widely applied to the field of biomedicine, and one important application is a molecular diagnosis technology based on the microfluidic chip.
Nucleic acid is a molecular biomarker widely applied to aspects of molecular diagnosis, food safety control, environmental monitoring and the like. Nucleic acid detection, which mainly involves extraction, amplification, detection steps, is a labor intensive, expensive, time consuming and largely instrument dependent method. With the increasing incidence of infectious diseases, food and water borne diseases, instant detection is becoming a focus of more and more attention.
Clinically, molecular diagnostics is widely used for the detection of infectious diseases, tumors, and genetic diseases. With the breakthrough of life science, biotechnology and micromachining technology, the related industries of high-end nucleic acid molecule detection become new global future competitive fields.
Molecular diagnosis is a fine detection method involving multiple amplification technologies, wherein a real-time isothermal fluorescence nucleic acid amplification and detection technology (SAT for short) is a novel nucleic acid detection technology combining a new generation of isothermal nucleic acid amplification technology and a real-time fluorescence detection technology. The technology has the advantages of rapid reaction, high sensitivity, high specificity, low pollution, stable reaction and the like. However, like the conventional nucleic acid detection technology, the existing detection device using the technology does not get rid of the disadvantages of long sample preparation time, complicated operation and the like, and does not meet the requirements of quick and low-cost clinical diagnosis.
Disclosure of Invention
The main object of the present invention is to overcome at least one of the above drawbacks and to provide a microfluidic chip, system and method for integrated nucleic acid extraction, amplification and detection.
In order to achieve the purpose, the invention adopts the following technical scheme:
the utility model provides a micro-fluidic chip of integrated nucleic acid extraction, amplification and detection, includes introduction port, washing liquid storage chamber, eluent storage chamber, nucleic acid extraction chamber, washing waste liquid receiving part and constant temperature amplification chamber, the introduction port is connected the nucleic acid extraction chamber, the nucleic acid extraction chamber is provided with the filter paper that possesses the nucleic acid extraction function, washing liquid storage chamber with eluent storage chamber respectively through the runner that is provided with the check valve with nucleic acid extraction chamber links to each other, nucleic acid extraction chamber connects washing waste liquid receiving part with constant temperature amplification chamber, washing waste liquid receiving part with constant temperature amplification chamber connects respectively and accesss to the controllable hole of external switching, wherein, washing liquid prestore of washing liquid in washing liquid storage chamber, eluent in eluent storage chamber, washing liquid storage chamber is used for prestore washing liquid under the external force drive and breaks through the check valve flow path the nucleic acid extraction chamber washes, the waste liquid flows to the washing waste liquid receiving part, the eluent storage cavity is used for enabling the eluent to break through the one-way valve to enter the nucleic acid extraction cavity for elution under the driving of external force, the eluted nucleic acid solution flows into the constant-temperature amplification cavity, and the constant-temperature amplification cavity is used for carrying out nucleic acid constant-temperature amplification and is provided with a transparent area.
Further:
the micro-fluidic chip comprises a base layer and a working layer which are stacked, wherein the sample inlet is provided with a washing liquid storage cavity, the eluent storage cavity is provided with a nucleic acid extraction cavity, the washing waste liquid receiving part and the constant-temperature amplification cavity are arranged on the working layer.
Washing waste liquid receiving unit includes washing waste liquid chamber, the controllable hole of switching includes washing waste liquid chamber exhaust hole and constant temperature amplification chamber exhaust hole, the nucleic acid draws the chamber and connects washing waste liquid chamber, washing waste liquid chamber exhaust hole is connected to washing waste liquid chamber, constant temperature amplification chamber connects constant temperature amplification chamber exhaust hole washing waste liquid chamber exhaust hole is opened and under the state that constant temperature amplification chamber exhaust hole closed, washing waste liquid flows in washing waste liquid chamber the washing waste liquid chamber exhaust hole is closed and under the state that constant temperature amplification chamber exhaust hole was opened, the eluant flows in constant temperature amplification chamber.
The micro-fluidic chip also comprises an elution waste liquid cavity, the nucleic acid extraction cavity is connected with the elution waste liquid cavity, and elution waste liquid flows into the elution waste liquid cavity.
The upper portion of the wash solution storage chamber and/or the eluent storage chamber is closed by a flexible membrane, and the stored liquid is released by applying external pressure to the flexible membrane.
The base layer and the working layer are made of one or more of polyvinyl chloride, polyethylene, polypropylene, polystyrene, polycarbonate or ABS, and are packaged together through a pressing process.
The microfluidic chip comprises a plurality of constant-temperature amplification cavities which are arranged in parallel.
A system for integrating nucleic acid extraction, amplification and detection comprises the centrifugal microfluidic chip.
A real-time fluorescent nucleic acid amplification detection method is used for carrying out real-time fluorescent nucleic acid amplification detection by using the microfluidic chip, and comprises the following steps:
(1) injecting a sample to be detected into the nucleic acid extraction cavity from the sample inlet;
(2) opening a hole connected with the washing waste liquid receiving part, closing a hole connected with the constant-temperature amplification cavity, pressurizing the washing liquid storage cavity, enabling the washing liquid to flow through the nucleic acid extraction cavity for washing, and enabling waste liquid to flow to the washing waste liquid receiving part;
(3) closing the hole connected with the washing waste liquid receiving part, opening the hole connected with the constant-temperature amplification cavity, pressurizing the eluent storage cavity, enabling eluent to slowly flow through the nucleic acid extraction cavity, and enabling the eluted nucleic acid solution to flow to the constant-temperature amplification cavity;
(4) controlling the temperature of the constant-temperature amplification cavity to be 37 ℃, observing the fluorescence intensity of the reactant under a microscope, and quantitatively analyzing the concentration of the nucleic acid.
Further, the isothermal amplification chamber maintains the reaction condition of 37 ℃ by contacting an external isothermal control plate.
The invention has the following beneficial effects:
the invention provides a micro-fluidic chip, a system and a method for integrating nucleic acid extraction, amplification and detection, which are used for extracting nucleic acid, performing constant-temperature amplification and performing real-time fluorescence detection. The invention realizes the extraction, amplification and detection of integrated nucleic acid on a microfluidic chip, can realize the automatic adsorption, extraction, purification, amplification and detection of nucleic acid on the chip, and shortens the amplification and detection time. The chip can be processed with low cost, produced in batch and convenient to store and transport. All reagents required by the reaction are pre-embedded in the chip, manual addition is not needed, operation steps are reduced, meanwhile, one chip can detect multiple indexes, detection paths are independent and cannot influence each other, and the reliable, stable and quick obtaining of detection results is sequentially guaranteed. The invention can reduce the complex instruments and reagents used in the processes of nucleic acid extraction, amplification and detection, effectively reduce the detection cost and improve the detection efficiency; in addition, the chip can be used as a disposable product, and the sample inlet is sealed after sample adding, so that potential positive pollution can be effectively avoided.
According to the invention, real-time fluorescent nucleic acid constant-temperature amplification and detection are realized on the microfluidic chip by the technical scheme, and the nucleic acid extraction process is integrated on the chip, so that the complicated manual operation is simplified, the advantages of low cost, low reagent consumption, high flux, automatic process and the like of the microfluidic chip technology can be fully exerted, the advantages of rapidness, simplicity, stability, reliability and the like of the fluorescent nucleic acid constant-temperature amplification and detection technology are fully exerted, and the requirements of clinical and epidemic disease detection and other rapid detection are met. The chip can be widely used in the field of rapid diagnosis of nucleic acid molecules, reduces the detection cost and improves the detection efficiency.
Drawings
FIG. 1 is a front perspective structural view of a working layer of a microfluidic chip integrating nucleic acid extraction, amplification and real-time fluorescence detection according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of a reverse perspective view of a working layer of a microfluidic chip integrating nucleic acid extraction, amplification and real-time fluorescence detection according to an embodiment of the present invention.
Detailed Description
The embodiments of the present invention will be described in detail below. It should be emphasized that the following description is merely exemplary in nature and is not intended to limit the scope of the invention or its application.
It will be understood that when an element is referred to as being "secured to" or "disposed on" another element, it can be directly on the other element or be indirectly on the other element. When an element is referred to as being "connected to" another element, it can be directly connected to the other element or be indirectly connected to the other element. In addition, the connection may be for either a fixed function or a circuit/signal communication function.
It is to be understood that the terms "length," "width," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like are used in an orientation or positional relationship indicated in the drawings for convenience in describing the embodiments of the present invention and to simplify the description, and are not intended to indicate or imply that the referenced device or element must have a particular orientation, be constructed in a particular orientation, and be in any way limiting of the present invention.
Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the embodiments of the present invention, "a plurality" means two or more unless specifically limited otherwise.
Referring to fig. 1 and fig. 2, an embodiment of the present invention provides a microfluidic chip integrating nucleic acid extraction, amplification and detection, which has a sample inlet 4, a washing solution storage chamber 3, an eluent storage chamber 11, a nucleic acid extraction chamber 10, a washing waste liquid receiving portion and a constant temperature amplification chamber 10, wherein the sample inlet 4 is connected to the nucleic acid extraction chamber 10, the nucleic acid extraction chamber 10 is provided with filter paper having a nucleic acid extraction function, preferably, a washing solution inlet 2 and an eluent inlet 1 are further provided, the washing solution inlet 2 is connected to the washing solution storage chamber 3, the eluent inlet 1 is connected to the eluent storage chamber 11, the washing solution storage chamber 3 and the eluent storage chamber 11 are respectively connected to the nucleic acid extraction chamber 10 through a flow channel provided with a check valve, the nucleic acid extraction chamber 10 is connected to the washing waste liquid receiving portion and the constant temperature amplification chamber 10, the washing waste liquid receiving part and the constant temperature amplification chamber 10 are respectively connected with a hole (such as a waste liquid chamber exhaust hole 5 and a constant temperature amplification chamber exhaust hole 7) which is opened and closed and is controlled to the outside, wherein, the washing liquid storage cavity 3 is pre-stored with washing liquid, the eluent storage cavity 11 is pre-stored with eluent, the washing liquid storage cavity 3 is used for enabling the washing liquid to break through the one-way valve and flow through the nucleic acid extraction cavity 10 for washing under the driving of external force, the waste liquid flows to the washing waste liquid receiving part, the eluent storage cavity 11 is used for enabling the eluent to break through the one-way valve to enter the nucleic acid extraction cavity 10 for elution under the driving of external force, the eluted nucleic acid solution flows into the constant-temperature amplification cavity 10, the constant-temperature amplification cavity 10 is provided with a specific primer and DNA polymerase, for mixing with a nucleic acid solution to perform isothermal amplification of nucleic acids, the isothermal amplification chamber 10 has a transparent region for detection. Wherein the controllable opening and closing hole can be controlled by using a matched sealing plug.
The micro-fluidic chip integrating nucleic acid extraction, amplification and detection provided by the embodiment of the invention is used for extracting nucleic acid, performing constant-temperature amplification and performing real-time fluorescence detection. The embodiment of the invention realizes the extraction, amplification and detection of integrated nucleic acid on one microfluidic chip, can realize automatic nucleic acid adsorption, extraction, purification, amplification and detection on the chip, and shortens the amplification and detection time. The chip can be processed with low cost, produced in batch and convenient to store and transport. All reagents required by the reaction are pre-embedded in the chip, manual addition is not needed, operation steps are reduced, meanwhile, one chip can detect multiple indexes, detection paths are independent and cannot influence each other, and the reliable, stable and quick obtaining of detection results is sequentially guaranteed. The invention can reduce the complex instruments and reagents used in the processes of nucleic acid extraction, amplification and detection, effectively reduce the detection cost and improve the detection efficiency; in addition, the chip can be used as a disposable product, and the sample inlet 4 is sealed after sample adding, so that potential positive pollution can be effectively avoided.
In the preferred embodiment, the micro-fluidic chip includes stratum basale and the working layer of range upon range of setting, the introduction port 4, the washing liquid inlet 2, eluent inlet 1, washing liquid storage chamber 3, eluent storage chamber 11, nucleic acid extraction chamber 10, washing waste liquid receiving part, constant temperature amplification chamber 10 and corresponding runner set up on the working layer.
In a preferred embodiment, the washing waste liquid receiving unit includes a washing waste liquid chamber 6, the opening/closing controllable hole includes a washing waste liquid chamber vent hole 5 and a constant temperature amplification chamber vent hole 7, the nucleic acid extraction chamber 10 is connected to the washing waste liquid chamber 6, the washing waste liquid chamber 6 is connected to the washing waste liquid chamber vent hole 5, the constant temperature amplification chamber 10 is connected to the constant temperature amplification chamber vent hole 7, the washing waste liquid flows into the washing waste liquid chamber 6 in a state where the washing waste liquid chamber vent hole 5 is opened and the constant temperature amplification chamber vent hole 7 is closed, and the eluent flows into the constant temperature amplification chamber 10 in a state where the washing waste liquid chamber vent hole 5 is closed and the constant temperature amplification chamber vent hole 7 is opened.
In a preferred embodiment, the microfluidic chip further comprises an elution waste liquid cavity 9, the nucleic acid extraction cavity 10 is connected with the elution waste liquid cavity 9, and elution waste liquid flows into the elution waste liquid cavity 9.
In a preferred embodiment, the upper portion of the wash solution storage chamber 3 and/or the eluent storage chamber 11 is closed by a flexible membrane, and the stored liquid is released by applying external pressure to the flexible membrane.
In some embodiments, the material of the substrate layer and the working layer comprises one or more of polyvinyl chloride, polyethylene, polypropylene, polystyrene, polycarbonate, or ABS.
In some embodiments, the substrate layer and the working layer are encapsulated together by a lamination process.
In a preferred embodiment, the microfluidic chip comprises a plurality of isothermal amplification chambers 10 arranged in parallel, so that a plurality of detections can be performed simultaneously.
The embodiment of the invention also provides a system for integrating nucleic acid extraction, amplification and detection, which comprises the centrifugal microfluidic chip.
The embodiment of the invention also provides a real-time fluorescent nucleic acid amplification detection method, which is used for carrying out real-time fluorescent nucleic acid amplification detection by using the microfluidic chip and comprises the following steps:
(1) injecting a sample to be detected into the nucleic acid extraction cavity 10 from the sample inlet 4;
(2) opening a hole connected with the washing waste liquid receiving part, closing a hole connected with the constant temperature amplification chamber 10, pressurizing the washing liquid storage chamber 3, allowing the washing liquid to flow through the nucleic acid extraction chamber 10 for washing, and allowing the waste liquid to flow to the washing waste liquid receiving part;
(3) closing the hole connected with the washing waste liquid receiving part, opening the hole connected with the constant-temperature amplification cavity 10, pressurizing the eluent storage cavity 11, enabling eluent to slowly flow through the nucleic acid extraction cavity 10, and enabling the eluted nucleic acid solution to flow to the constant-temperature amplification cavity 10;
(4) the temperature of the isothermal amplification chamber 10 was controlled to 37 ℃, the fluorescence intensity of the reactants was observed under a microscope, and the nucleic acid concentration was quantitatively analyzed.
In a preferred embodiment, the isothermal amplification chamber 10 maintains 37 ℃ reaction conditions by contacting an external isothermal control plate.
Specific embodiments of the present invention are further described below with reference to the accompanying drawings.
In one embodiment, the microfluidic chip for nucleic acid extraction, amplification and detection can be functionally divided into a nucleic acid extraction unit and a nucleic acid isothermal amplification and detection unit; the chip body is provided with a sample inlet 4, a nucleic acid extraction cavity 10, a washing liquid storage cavity 3, an eluent storage cavity 11, a constant temperature amplification cavity 10, a washing waste liquid cavity 6 and an elution waste liquid cavity 9. The washing liquid stores up chamber 3 and the eluent and stores up and has had required reagent solution in advance in the chamber 11 on the chip, through the runner intercommunication between each cavity, is equipped with the check valve in the washing liquid stores up chamber 3 and the eluent and stores up the exit flow way of chamber 11, but washing liquid stores up chamber 3 and eluent and stores up 11 plus drive in chamber. The chip can be used as a carrier for realizing nucleic acid adsorption, extraction, purification, amplification and detection after a sample is manually added, and realizes the functions of integrating the extraction, amplification and detection of nucleic acid on the chip. The chip can be a disposable product, and the sample inlet 4 is sealed after sample adding, so that potential positive pollution can be effectively avoided.
One particular chip structure includes a base layer and an active layer. The base layer and the working layer are made of common medical plastics such as polyvinyl chloride (PVC), Polyethylene (PE), polypropylene (PP), Polystyrene (PS), Polycarbonate (PC), ABS and the like, and are molded by various plastic molding modes such as mold pressing thermoplastic molding, injection molding and the like. The substrate layer and the working layer may be encapsulated together using a press-fit process.
Wherein, the chip structure contains sample inlet 4, nucleic acid extraction chamber 10, washing liquid storage chamber 3, eluent storage chamber 11, constant temperature amplification chamber 10 (be reaction chamber), washing waste liquid chamber 6 and elution waste liquid chamber 9, communicates through the runner between the chamber. A filter paper (e.g., Whatman No.1 filter paper) having a nucleic acid extraction function is fixed in advance in the nucleic acid extraction chamber 10. The isothermal amplification cavity 10 is internally provided with a specific primer and DNA polymerase.
Wherein, the accessible the artifical injection sample of introduction port 4, and the accessible drive washing liquid storage chamber 3 with washing liquid and the eluant of prestoring in the eluant storage chamber 11 lets in proper order in nucleic acid extraction chamber 10 and wait to extract nucleic acid sample, washing liquid, eluant etc. and the waste liquid flows into the waste liquid chamber, and the nucleic acid solution under the elution flows into nucleic acid constant temperature amplification and detecting element.
Wherein, be provided with the check valve in advance in the intercommunication outlet flow channel with washing liquid storage chamber 3 and eluant storage chamber 11 to realize the stable storage of washing liquid and eluant. After the washing liquid storage cavity 3 and the eluent storage cavity 11 are pressurized by external drive, liquid in the cavities is released from the cavities and breaks through the one-way valve to enter the nucleic acid extraction cavity 10. The eluted nucleic acid solution enters the constant temperature amplification cavity 10 and dissolves the specific primer and DNA polymerase which are freeze-dried in advance in the cavity to form a mixed solution, and the nucleic acid is amplified at constant temperature. The temperature of the constant temperature amplification cavity 10 is maintained at 37 ℃ by a constant temperature control plate which is contacted with the outside, and the cyclic amplification condition is reflected by placing the chip in a fluorescence detection system and detecting the change of fluorescence intensity in real time.
The nucleic acid detection method of the embodiment of the invention is based on a real-time fluorescent nucleic acid amplification detection technology. The nucleic acid detection method of an embodiment may include the steps of:
injecting a sample to be detected into the nucleic acid extraction cavity 10 to the sample inlet 4 of the nucleic acid extraction unit by using an injector, wherein DNA is adsorbed by the filter paper, and the rest waste liquid flows to a waste liquid cavity;
pressurizing the washing liquid storage cavity 3, washing the washing liquid by flowing through the nucleic acid extraction cavity 10, and flowing the waste liquid to the waste liquid cavity;
closing the waste liquid cavity exhaust hole 5 and opening the reaction cavity exhaust hole;
pressurizing the eluent storage cavity 11, slowly flowing eluent through the nucleic acid extraction cavity 10, and flowing the eluted nucleic acid solution to the reaction cavity;
controlling the temperature of the nucleic acid isothermal amplification and detection unit to be 37 ℃, observing the fluorescence intensity of the reactant under a microscope, and quantitatively analyzing the concentration of the nucleic acid.
The structure of the microfluidic chip integrating nucleic acid extraction, amplification and real-time fluorescence detection in one embodiment is shown in fig. 1-2, and the working layer of the chip comprises an eluent liquid inlet 1, a washing liquid inlet 2, a washing liquid storage cavity 3, a sample inlet 4, a washing waste liquid cavity exhaust hole 5, a washing waste liquid cavity 6, a constant temperature amplification cavity exhaust hole 7, a constant temperature amplification cavity 8, an elution waste liquid cavity 9, a nucleic acid extraction cavity 10 and a plurality of flow channels which are communicated with one another.
The material of the working layer can adopt common medical plastics, such as polyvinyl chloride (PVC), Polyethylene (PE), polypropylene (PP), Polystyrene (PS), Polycarbonate (PC) and ABS, and adopts various plastic forming modes such as mould pressing thermoplastic forming, injection molding and the like; for example, an injection molding method is adopted to process a mold in advance, then a polypropylene (PP) material is melted in a constant temperature cylinder (20-280 ℃), then the melted PP material is injected into the mold under pressure (800-. Before assembling the chip, the primers and the DNA polymerase need to be freeze-dried in the constant temperature amplification cavity 8 in advance, filter paper with a nucleic acid extraction function needs to be fixedly inlaid in the nucleic acid extraction cavity 10, and then the working layer is plastically packaged by a flexible thin film. The sealing plugs of the exhaust holes and the sampling holes have the same specification, and are also made of medical plastics which are the same as the working layer. After the chip is assembled, the eluent is required to be quantitatively injected through an eluent inlet 1; after quantitative injection of the washing solution through the washing solution inlet 2, the eluent inlet 1, the washing solution inlet 2 and the constant-temperature amplification cavity exhaust hole 7 are sealed by sealing plugs.
The steps of nucleic acid extraction, amplification and detection of one embodiment comprise:
1. injecting a sample to be detected to the nucleic acid extraction cavity 10 from the sample inlet 4 by using an injector, and adsorbing DNA in the sample by the filter paper;
2. a pressurizing device is used for slowly applying pressure to the flexible film above the washing liquid storage cavity 3, the washing liquid flows through the filter paper in the nucleic acid extraction cavity 10 to fully wash the DNA adsorbed on the filter paper, and the washed waste liquid flows to the washing waste liquid cavity 6;
3. taking down the sealing plug on the exhaust hole 7 of the constant-temperature amplification cavity, and sealing the exhaust hole 5 of the washing waste liquid cavity by using the sealing plug;
4. a pressurizing device is used for slowly applying pressure to the flexible film above the eluent storage cavity 11, the eluent flows through the filter paper in the nucleic acid extraction cavity 10 and elutes the DNA adsorbed on the filter paper, and the eluent containing the DNA uniformly flows to each constant temperature amplification cavity 8;
5. the primers and the enzyme which are pre-frozen and dried in the constant temperature amplification cavity 8 are fully dissolved by the eluent containing the DNA;
6. controlling the temperature of the nucleic acid isothermal amplification and detection unit to be 37 ℃, observing the fluorescence intensity of the reactant under a microscope, and quantitatively analyzing the concentration of the nucleic acid.
According to the invention, real-time fluorescent nucleic acid constant-temperature amplification and detection are realized on the microfluidic chip by the technical scheme, and the nucleic acid extraction process is integrated on the chip, so that the complicated manual operation is simplified, the advantages of low cost, low reagent consumption, high flux, automatic process and the like of the microfluidic chip technology can be fully exerted, the advantages of rapidness, simplicity, stability, reliability and the like of the fluorescent nucleic acid constant-temperature amplification and detection technology are fully exerted, and the requirements of clinical and epidemic disease detection and other rapid detection are met. The chip can be widely used in the field of rapid diagnosis of nucleic acid molecules, reduces the detection cost and improves the detection efficiency.
The background of the present invention may contain background information related to the problem or environment of the present invention and does not necessarily describe the prior art. Accordingly, the inclusion in the background section is not an admission of prior art by the applicant.
The foregoing is a more detailed description of the invention in connection with specific/preferred embodiments and is not intended to limit the practice of the invention to those descriptions. It will be apparent to those skilled in the art that various substitutions and modifications can be made to the described embodiments without departing from the spirit of the invention, and these substitutions and modifications should be considered to fall within the scope of the invention. In the description herein, references to the description of the term "one embodiment," "some embodiments," "preferred embodiments," "an example," "a specific example," or "some examples" or the like are intended to mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Various embodiments or examples and features of various embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction. Although embodiments of the present invention and their advantages have been described in detail, it should be understood that various changes, substitutions and alterations can be made herein without departing from the scope of the claims.
Claims (10)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911328740.7A CN111073810A (en) | 2019-12-20 | 2019-12-20 | Microfluidic chip, system and method integrating nucleic acid extraction, amplification and detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911328740.7A CN111073810A (en) | 2019-12-20 | 2019-12-20 | Microfluidic chip, system and method integrating nucleic acid extraction, amplification and detection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111073810A true CN111073810A (en) | 2020-04-28 |
Family
ID=70316372
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911328740.7A Pending CN111073810A (en) | 2019-12-20 | 2019-12-20 | Microfluidic chip, system and method integrating nucleic acid extraction, amplification and detection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111073810A (en) |
Cited By (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111534428A (en) * | 2020-05-22 | 2020-08-14 | 中国科学院上海技术物理研究所 | Integrated nucleic acid extraction and amplification detection system based on PCR amplification |
| CN111575176A (en) * | 2020-05-22 | 2020-08-25 | 中国科学院上海技术物理研究所 | Closed automatic nucleic acid extraction and detection system based on CRISPR technology |
| CN111760601A (en) * | 2020-07-03 | 2020-10-13 | 中国科学院合肥物质科学研究院 | Microfluidic chip with integrated liquid circuit switching valve and nucleic acid detection method |
| CN112501258A (en) * | 2020-11-10 | 2021-03-16 | 清华大学深圳国际研究生院 | Micro-fluidic chip for nucleic acid extraction, purification, amplification and detection |
| CN113337577A (en) * | 2021-05-31 | 2021-09-03 | 深圳市博德致远生物技术有限公司 | Application method of micro-fluidic chip |
| CN113684118A (en) * | 2021-07-14 | 2021-11-23 | 浙江大学 | Integrated nucleic acid analysis chip |
| CN113846014A (en) * | 2021-10-29 | 2021-12-28 | 青岛全诊生物技术有限公司 | Nucleic acid extraction and amplification device |
| CN113913289A (en) * | 2021-12-07 | 2022-01-11 | 北京芯源视界生物科技有限公司 | Detection kit and detection method for pathogenic nucleic acid under airtight condition |
| CN114058088A (en) * | 2020-07-29 | 2022-02-18 | 京东方科技集团股份有限公司 | Gel precursor, nucleic acid amplification reagent gel, memory chip and using method |
| CN114875121A (en) * | 2022-06-23 | 2022-08-09 | 浙江大学 | Device and method for detecting nucleic acid amplification product by adopting lateral chromatography test strip |
| CN115074207A (en) * | 2021-03-11 | 2022-09-20 | 上海交通大学 | Microfluidic device for biological samples and nucleic acid extraction and amplification method |
| CN115232728A (en) * | 2022-08-05 | 2022-10-25 | 中国水稻研究所 | Molecular detection method and device suitable for field detection |
| CN115305190A (en) * | 2022-08-05 | 2022-11-08 | 中国水稻研究所 | High-throughput detection device based on microfluidics and nucleic acid amplification |
| CN115449478A (en) * | 2022-10-14 | 2022-12-09 | 上海晶锴科技有限公司 | A reaction plate and a device for parallel determination of multiple nucleic acid molecule samples |
| CN115725399A (en) * | 2022-11-29 | 2023-03-03 | 深圳市儿童医院 | Microfluidic chip and application thereof |
| CN115896245A (en) * | 2022-12-29 | 2023-04-04 | 币冠(上海)生物科技有限公司 | A rapid detection method for fully automatic nucleic acid extraction and multiplex polymerase chain reaction based on microfluidic chip |
| WO2023168760A1 (en) * | 2022-03-10 | 2023-09-14 | 广东和信健康科技有限公司 | Detection consumable and detection apparatus |
| CN118146933A (en) * | 2024-05-13 | 2024-06-07 | 至美时代生物智能科技(北京)有限公司 | Microorganism detection device, detection system and detection method |
| CN118384935A (en) * | 2024-04-24 | 2024-07-26 | 深圳市呈晖医疗科技有限公司 | Check valve structure on centrifugal micro-fluidic chip |
| CN119464008A (en) * | 2024-11-22 | 2025-02-18 | 上海交通大学 | Whole blood sample nucleic acid extraction cartridge |
| CN120644259A (en) * | 2025-06-18 | 2025-09-16 | 中国科学院半导体研究所 | Microfluidic chip and sample detection method |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007106580A2 (en) * | 2006-03-15 | 2007-09-20 | Micronics, Inc. | Rapid magnetic flow assays |
| CN105349401A (en) * | 2015-10-14 | 2016-02-24 | 安徽易康达光电科技有限公司 | Multifunctional integrated microfluidic nucleic acid analysis chip and preparation and analysis method thereof |
| CN106591104A (en) * | 2009-07-24 | 2017-04-26 | 阿科尼生物系统公司 | Flow cell device |
| CN206375901U (en) * | 2016-12-14 | 2017-08-04 | 深圳市埃克特生物科技有限公司 | A kind of integrated closed microfluidic nucleic acid detection means |
| CN108865658A (en) * | 2018-08-27 | 2018-11-23 | 苏州绘真医学检验有限公司 | A kind of tubular type plasma free method for extracting nucleic acid and system |
| CN109207475A (en) * | 2018-10-22 | 2019-01-15 | 博迪泰（厦门）生物科技有限公司 | A kind of Rapid nucleic acid extracting method |
| CN110142066A (en) * | 2019-04-25 | 2019-08-20 | 深圳市刚竹医疗科技有限公司 | Micro-fluidic chip and analysis system |
| CN110452806A (en) * | 2018-05-08 | 2019-11-15 | 国家纳米科学中心 | Fully automated microfluidic nucleic acid detection chip based on loop-mediated isothermal amplification, its kit and method |
-
2019
- 2019-12-20 CN CN201911328740.7A patent/CN111073810A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007106580A2 (en) * | 2006-03-15 | 2007-09-20 | Micronics, Inc. | Rapid magnetic flow assays |
| CN106591104A (en) * | 2009-07-24 | 2017-04-26 | 阿科尼生物系统公司 | Flow cell device |
| CN105349401A (en) * | 2015-10-14 | 2016-02-24 | 安徽易康达光电科技有限公司 | Multifunctional integrated microfluidic nucleic acid analysis chip and preparation and analysis method thereof |
| CN206375901U (en) * | 2016-12-14 | 2017-08-04 | 深圳市埃克特生物科技有限公司 | A kind of integrated closed microfluidic nucleic acid detection means |
| CN110452806A (en) * | 2018-05-08 | 2019-11-15 | 国家纳米科学中心 | Fully automated microfluidic nucleic acid detection chip based on loop-mediated isothermal amplification, its kit and method |
| CN108865658A (en) * | 2018-08-27 | 2018-11-23 | 苏州绘真医学检验有限公司 | A kind of tubular type plasma free method for extracting nucleic acid and system |
| CN109207475A (en) * | 2018-10-22 | 2019-01-15 | 博迪泰（厦门）生物科技有限公司 | A kind of Rapid nucleic acid extracting method |
| CN110142066A (en) * | 2019-04-25 | 2019-08-20 | 深圳市刚竹医疗科技有限公司 | Micro-fluidic chip and analysis system |
Cited By (25)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111575176A (en) * | 2020-05-22 | 2020-08-25 | 中国科学院上海技术物理研究所 | Closed automatic nucleic acid extraction and detection system based on CRISPR technology |
| CN111534428A (en) * | 2020-05-22 | 2020-08-14 | 中国科学院上海技术物理研究所 | Integrated nucleic acid extraction and amplification detection system based on PCR amplification |
| CN111760601B (en) * | 2020-07-03 | 2022-04-22 | 中国科学院合肥物质科学研究院 | Microfluidic chip with integrated liquid circuit switching valve and nucleic acid detection method |
| CN111760601A (en) * | 2020-07-03 | 2020-10-13 | 中国科学院合肥物质科学研究院 | Microfluidic chip with integrated liquid circuit switching valve and nucleic acid detection method |
| CN114058088A (en) * | 2020-07-29 | 2022-02-18 | 京东方科技集团股份有限公司 | Gel precursor, nucleic acid amplification reagent gel, memory chip and using method |
| CN112501258A (en) * | 2020-11-10 | 2021-03-16 | 清华大学深圳国际研究生院 | Micro-fluidic chip for nucleic acid extraction, purification, amplification and detection |
| CN115074207A (en) * | 2021-03-11 | 2022-09-20 | 上海交通大学 | Microfluidic device for biological samples and nucleic acid extraction and amplification method |
| CN113337577A (en) * | 2021-05-31 | 2021-09-03 | 深圳市博德致远生物技术有限公司 | Application method of micro-fluidic chip |
| CN113684118A (en) * | 2021-07-14 | 2021-11-23 | 浙江大学 | Integrated nucleic acid analysis chip |
| CN113684118B (en) * | 2021-07-14 | 2024-05-28 | 浙江大学 | An integrated nucleic acid analysis chip |
| CN113846014A (en) * | 2021-10-29 | 2021-12-28 | 青岛全诊生物技术有限公司 | Nucleic acid extraction and amplification device |
| CN113913289B (en) * | 2021-12-07 | 2022-03-22 | 北京芯源视界生物科技有限公司 | Detection kit and detection method for pathogenic nucleic acid under airtight condition |
| CN113913289A (en) * | 2021-12-07 | 2022-01-11 | 北京芯源视界生物科技有限公司 | Detection kit and detection method for pathogenic nucleic acid under airtight condition |
| WO2023168760A1 (en) * | 2022-03-10 | 2023-09-14 | 广东和信健康科技有限公司 | Detection consumable and detection apparatus |
| CN114875121B (en) * | 2022-06-23 | 2026-01-30 | 浙江大学 | Apparatus and method for detecting nucleic acid amplification products using lateral chromatography test strips |
| CN114875121A (en) * | 2022-06-23 | 2022-08-09 | 浙江大学 | Device and method for detecting nucleic acid amplification product by adopting lateral chromatography test strip |
| CN115232728A (en) * | 2022-08-05 | 2022-10-25 | 中国水稻研究所 | Molecular detection method and device suitable for field detection |
| CN115305190A (en) * | 2022-08-05 | 2022-11-08 | 中国水稻研究所 | High-throughput detection device based on microfluidics and nucleic acid amplification |
| CN115449478A (en) * | 2022-10-14 | 2022-12-09 | 上海晶锴科技有限公司 | A reaction plate and a device for parallel determination of multiple nucleic acid molecule samples |
| CN115725399A (en) * | 2022-11-29 | 2023-03-03 | 深圳市儿童医院 | Microfluidic chip and application thereof |
| CN115896245A (en) * | 2022-12-29 | 2023-04-04 | 币冠(上海)生物科技有限公司 | A rapid detection method for fully automatic nucleic acid extraction and multiplex polymerase chain reaction based on microfluidic chip |
| CN118384935A (en) * | 2024-04-24 | 2024-07-26 | 深圳市呈晖医疗科技有限公司 | Check valve structure on centrifugal micro-fluidic chip |
| CN118146933A (en) * | 2024-05-13 | 2024-06-07 | 至美时代生物智能科技(北京)有限公司 | Microorganism detection device, detection system and detection method |
| CN119464008A (en) * | 2024-11-22 | 2025-02-18 | 上海交通大学 | Whole blood sample nucleic acid extraction cartridge |
| CN120644259A (en) * | 2025-06-18 | 2025-09-16 | 中国科学院半导体研究所 | Microfluidic chip and sample detection method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111073810A (en) | Microfluidic chip, system and method integrating nucleic acid extraction, amplification and detection | |
| CN111088161B (en) | A microfluidic chip and detection method for nucleic acid amplification detection | |
| US12275009B2 (en) | Microfluidic cartridge | |
| CN211921576U (en) | Micro-fluidic chip and system integrating nucleic acid extraction, amplification and detection | |
| CN111073811B (en) | A microfluidic chip and detection method for real-time fluorescent nucleic acid amplification detection | |
| KR101984699B1 (en) | Micro-fluidic system for analysis of nucleic acid | |
| US9250169B2 (en) | Selective capture and release of analytes | |
| US20090130658A1 (en) | Arrangement for integrated and automated dna or protein analysis in a single-use cartridge, method for producing such a cartridge and operating method for dna or protein analysis using such a cartridge | |
| US20050221281A1 (en) | Self-contained microfluidic biochip and apparatus | |
| CA3141275A1 (en) | Systems for sample analysis | |
| CN115814864B (en) | In vitro diagnostic analysis devices and reagent cartridges | |
| CN110114145B (en) | Analysis system and method for detecting sample | |
| CN113649095A (en) | A highly integrated microfluidic chip for nucleic acid detection and method of use | |
| CN210522535U (en) | Micro-fluidic chip | |
| CN113444624A (en) | Nucleic acid detection chip driven by tensile force and nucleic acid detection equipment | |
| CN117551540A (en) | Centrifugal microfluidic nucleic acid analysis chip and nucleic acid detection method | |
| JP2007071555A (en) | Substrate having protein immobilized thereon and microreactor using it | |
| CN211713056U (en) | Micro-fluidic chip for real-time fluorescent nucleic acid amplification detection | |
| JP2007136379A (en) | Micro-reactor and its manufacturing method | |
| CN115236346A (en) | Micro-fluidic detection chip | |
| JP4687413B2 (en) | Method for mixing two or more liquids in a microchip and a micro total analysis system | |
| TWI760373B (en) | Method and analysis system for testing a sample | |
| CN118146930A (en) | Single-disk full-extraction chip based on centrifugal microfluidic | |
| WO2007058077A1 (en) | Gene test method, microreactor for gene test and gene test system | |
| CN211755084U (en) | Micro-fluidic chip and can be sealed advance a kind structure |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200428 |
|
| RJ01 | Rejection of invention patent application after publication |