CN115491297A - Multilayer micro-fluidic chip integrating nucleic acid extraction, purification and digital detection - Google Patents

Multilayer micro-fluidic chip integrating nucleic acid extraction, purification and digital detection Download PDF

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Publication number
CN115491297A
CN115491297A CN202211031355.8A CN202211031355A CN115491297A CN 115491297 A CN115491297 A CN 115491297A CN 202211031355 A CN202211031355 A CN 202211031355A CN 115491297 A CN115491297 A CN 115491297A
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storage cavity
liquid storage
nucleic acid
detection
micro
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CN202211031355.8A
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Inventor
冯世伦
黄亚茹
高则航
贾春平
赵建龙
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Shanghai Institute of Microsystem and Information Technology of CAS
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Shanghai Institute of Microsystem and Information Technology of CAS
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    • 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
    • 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
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • 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
    • C12Q1/686—Polymerase chain reaction [PCR]
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention relates to a multilayer micro-fluidic chip integrating nucleic acid extraction, purification and digital detection, which comprises a PDMS-based nucleic acid extraction and purification layer (100), a glass substrate (200) and a silicon-based micro-cavity digital PCR detection layer (300) from top to bottom. The invention integrates sample preparation and digital PCR detection, and can be applied to accurate quantitative detection of pathogens; compared with the existing pathogen detection platform, the method has great advantages in the aspects of detection sensitivity, flow automation, integration and the like, and has wide application prospects in the aspect of molecular diagnosis.

Description

Multilayer micro-fluidic chip integrating nucleic acid extraction, purification and digital detection
Technical Field
The invention belongs to the field of pathogen detection platforms, and particularly relates to a multilayer microfluidic chip integrating nucleic acid extraction and purification and digital detection.
Background
The nucleic acid detection technology is the most widely applied molecular diagnosis technology in recent years, can quickly and efficiently amplify the characteristic nucleotide sequence of pathogenic bacteria in infectious disease diagnosis, and is widely applied in the fields of clinical diagnosis, disease screening and the like. Conventional nucleic acid detection procedures typically include: sample inactivation, nucleic acid extraction and purification, nucleic acid amplification, detection and the like. The extraction and purification of nucleic acid is the key in molecular biology experiments, and the relationship between the extraction and purification of nucleic acid with high quality can directly influence the success or failure of subsequent experiments. For nucleic acid extraction and purification, the reagents provided by the existing kit are often complicated in steps, long in time consumption and easy to cause cross contamination due to an operation mode or an experimental environment, so that the kit has high requirements on the experimental capability and the detection environment of medical personnel.
The nucleic acid amplification and detection is to perform high-efficiency and rapid amplification on the extracted and purified characteristic nucleotide sequence of the pathogenic bacteria and perform fluorescence or visual detection to obtain a real-time diagnosis result. The length, sensitivity and specificity of the nucleic acid amplification method will also directly affect the interpretation of subsequent results. The current method commonly used for nucleic acid amplification detection is real-time fluorescent quantitative PCR, which can perform qualitative and quantitative analysis on nucleic acid in a sample and is the gold standard of the current nucleic acid amplification detection. However, the method has disadvantages in that the quantitative analysis of the method must rely on the presence of a standard curve, absolute quantification of nucleic acid molecules cannot be achieved, and a target gene having low abundance cannot be detected when the nucleic acid content is low, resulting in false negative results.
In order to address the above-mentioned phenomena, research has been conducted to integrate nucleic acid extraction and purification and digital detection with higher sensitivity in a chip, and to simplify the detection process based on magnetic bead transfer or reagent flow distribution. The defects of the existing detection mode are that the integrated detection is difficult, the operation mode is complicated, and the liquid flow is often driven by means of negative pressure or centrifugation. Therefore, the application of a high-sensitivity and simple operation device integrating nucleic acid extraction and purification and digital detection in the field of molecular diagnosis still has the problem to be solved.
Disclosure of Invention
The invention aims to solve the technical problems that the prior art is difficult in integrated detection, complicated in operation mode and needs to drive liquid to flow by means of negative pressure or centrifugation.
The invention provides a multilayer micro-fluidic chip integrating nucleic acid extraction and purification and digital detection, which comprises a PDMS-based nucleic acid extraction and purification layer, a glass substrate and a silicon-based micro-cavity digital PCR detection layer from top to bottom; the PDMS-based nucleic acid extraction and purification layer consists of a liquid storage cavity, a bottom connecting pipeline, a micro-column array, a capillary explosion valve, a micro-cavity detection layer sample inlet connecting hole and a micro-cavity detection layer sample outlet connecting hole; the liquid storage cavity comprises a lysis solution storage cavity, a first washing solution storage cavity, a second washing solution storage cavity, a mineral oil storage cavity and an eluent liquid storage cavity which are sequentially connected through a bottom connecting pipeline; the micro-column array is used for water-oil separation of the mineral oil liquid storage cavity and the eluent liquid storage cavity, and the capillary explosion valve is used for preventing liquid in the eluent liquid storage cavity from spontaneously entering the sample inlet connecting hole in the nucleic acid purification process. And only when the mineral oil liquid storage cavity is filled with the oil phase to apply pressure, the water phase breaks through the capillary explosion valve and enters the silicon-based micro-cavity digital PCR detection layer through the sample inlet connecting hole.
The lysate liquid storage cavity, the first washing liquid storage cavity and the second washing liquid storage cavity realize the mutual noninterference of the lysate, the first washing liquid and the second washing liquid by using a water-in-oil principle.
The glass substrate is provided with openings corresponding to the sample inlet connecting hole of the microcavity detection layer and the sample outlet connecting hole of the microcavity detection layer.
The silicon-based micro-cavity digital PCR detection layer comprises a micro-cavity sample inlet, a flow channel, a micro-pore reaction chamber and a sample outlet; the microcavity sample inlet is connected with the microcavity detection layer sample inlet connecting hole, and the sample outlet is connected with the microcavity detection layer sample outlet connecting hole; the micropore reaction chamber is connected with the microcavity sample inlet and the microcavity sample outlet through a flow passage.
The eluent liquid storage cavity is provided with an eluent sample inlet hole.
The micropore reaction chamber is composed of a plurality of micropores distributed in the flow channel, the distance and the size of the micropore reaction chamber are kept consistent, and digital analysis of subsequent detection is facilitated.
The invention also provides a use method of the multilayer micro-fluidic chip integrating nucleic acid extraction and purification and digital detection, which comprises the following steps:
(1) Firstly, an aqueous elution reagent (water phase) is filled in an eluent liquid storage cavity in advance, then mineral oil is injected into a lysate liquid storage cavity, and the mineral oil flows through a first washing liquid storage cavity, a second washing liquid storage cavity and a mineral oil liquid storage cavity through a bottom connecting pipeline through an inclined chip and is filled; injecting nucleic acid extracting and purifying reagents into corresponding liquid storage cavities, wherein each reagent is separated into independent cavities;
(2) In the detection process, a sample is added into a lysis solution storage cavity, cells or viruses in the sample are lysed, released nucleic acid molecules are adsorbed by nucleic acid extraction magnetic beads, and then the nucleic acid extraction magnetic beads are dragged into a subsequent reaction chamber to complete the washing and elution processes, so that a PCR reaction solution is obtained;
(3) The adhesive tape seals other liquid storage cavities except the mineral oil liquid storage cavity, and the oil phase pushes the water phase to sequentially enter the silicon-based microcavity digital PCR detection layer through the microcavity detection layer sample inlet connecting hole by continuously injecting oil phase to apply pressure into the mineral oil liquid storage cavity; and finally, placing the chip into an in-situ PCR instrument, setting related programs, and observing the detection result under a fluorescence microscope after the reaction is finished.
The principle of the invention is as follows:
the invention integrates nucleic acid extraction and purification and digital PCR detection, designs a nucleic acid extraction and purification part based on a water and oil surface tension immiscible interface, and separates nucleic acid extraction and purification reagents in separate chambers through oil phase to form reagent droplets or maintains the water-oil interface through a micro-column array to enable the reagents to independently and stably exist in the chambers, thereby ensuring that the reagents independently react and do not interfere with each other in the operation process. The transfer of the nucleic acid molecules is completed by dragging the magnetic beads to shuttle in each reagent droplet through the bottom connecting channel so as to complete the adsorption, enrichment, washing and elution of the nucleic acid molecules. A digital PCR detection part is designed on the basis of preparing a sample by a nucleic acid extraction and purification part, and accurate absolute quantitative detection can be realized through limiting dilution and Poisson distribution statistics.
Advantageous effects
The multilayer microfluidic chip integrates sample preparation and digital PCR detection, and can be applied to accurate quantitative detection of pathogens. The specially designed water-oil separation structure (microcolumn array) has a reagent separation effect on a nucleic acid extraction and purification part, and when the subsequent microcavity digital detection is carried out, water and oil can enter the microcavity and be filled orderly only by continuously injecting oil phase into an oil cavity to provide pressure, and the liquid flow is driven by no complex equipment such as a negative pressure pump or a centrifugal machine, so that simple and convenient semi-automatic sample injection is realized. The invention can be sent into a PCR instrument to complete the detection of a target sample only by simply injecting a reagent and a magnet in the detection process, greatly reduces the requirements on the professional skills and auxiliary equipment of operators, and has potential applicability in the field of molecular diagnosis.
Drawings
FIG. 1 is a schematic structural diagram of a multilayer microfluidic chip according to the present invention;
FIG. 2 is a schematic structural diagram of a PDMS-based nucleic acid extraction and purification layer in a multi-layer microfluidic chip according to the present invention;
FIG. 3 is a schematic structural diagram of a silicon-based microcavity digital PCR detection layer in a multilayer microfluidic chip according to the present invention;
FIG. 4 is a cross-sectional view of a multilayer microfluidic chip of the present invention;
FIG. 5 is a schematic diagram of a process for using the multi-layer microfluidic chip of the present invention;
FIG. 6 shows the result of ORF1ab gene purification and digital detection of SARS-CoV-2 by using the multi-layer microfluidic chip of the present invention;
wherein (A) has a concentration of 10 5 cp/. Mu.L, (B) concentration 10 3 cp/. Mu.L, (C) concentration 10 1 cp/. Mu.L, blank (D).
The reference numerals in fig. 1 to 5 are as follows:
100 is PDMS-based nucleic acid extraction and purification layer;
101 is a bottom connecting pipeline;
102 is a micro-column array;
103 is an eluent sample inlet hole;
104 is a capillary explosion valve;
105 is a connection hole of a sample inlet of the microcavity detection layer;
106 is a sample outlet connecting hole of the microcavity detection layer;
110 is a liquid storage cavity;
111 is a lysate storage cavity;
112 is a first washing liquid storage cavity;
113 is a second washing liquid storage cavity;
114 is a mineral oil storage cavity;
115 is an eluent liquid storage cavity;
120 is a reaction reagent;
121 is lysate reagent;
122 is a first wash solution;
123 is a second wash solution;
124 is mineral oil;
125 elution (amplification) reagents;
131 is a nucleic acid extraction magnetic bead;
200 is a glass substrate;
300 is a silicon-based microcavity digital PCR detection layer;
301 is a microcavity sample inlet;
302 is a flow channel;
303 is a microporous reaction chamber;
and 304 is a sample outlet.
Detailed Description
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention can be made by those skilled in the art after reading the teaching of the present invention, and these equivalents also fall within the scope of the claims appended to the present application.
Example 1
As shown in fig. 1-2 and 4, the present embodiment provides a multi-layered microfluidic chip integrating nucleic acid extraction, purification and digital detection, which includes, from top to bottom, a PDMS-based nucleic acid extraction and purification layer 100, a glass substrate 200, and a silicon-based microcavity digital PCR detection layer 300; the PDMS-based nucleic acid extraction and purification layer 100 consists of a liquid storage cavity 110, a bottom connecting pipeline 101, a micro-column array 102, a capillary explosion valve 104, a micro-cavity detection layer sample inlet connecting hole 105 and a micro-cavity detection layer sample outlet connecting hole 106; the liquid storage cavity 110 comprises a lysis solution storage cavity 111, a first washing solution storage cavity 112, a second washing solution storage cavity 113, a mineral oil storage cavity 114 and an eluent liquid storage cavity 115 which are sequentially connected through a bottom connecting pipeline 101; the micro-column array 102 is used for water-oil separation of the mineral oil storage cavity 114 and the eluent storage cavity 115, the capillary blasting valve 104 is used for preventing liquid in the eluent storage cavity 115 from spontaneously entering the sample inlet connecting hole 105 in the nucleic acid purification process, and only when oil phase is injected into the mineral oil storage cavity 114 to apply pressure, the water phase can break through the capillary blasting valve 104 and enter the silicon-based micro-cavity digital PCR detection layer through the sample inlet connecting hole 105.
The lysate liquid storage cavity 111, the first washing liquid storage cavity 112 and the second washing liquid storage cavity 113 realize mutual noninterference of the lysate, the first washing liquid and the second washing liquid by using a water-in-oil principle.
The glass substrate 200 is provided with openings corresponding to the sample inlet connection hole 105 and the sample outlet connection hole 106 of the microcavity detection layer.
As shown in fig. 3, the silicon-based microcavity digital PCR detection layer 300 includes a microcavity sample inlet 301, a flow channel 302, a microporous reaction chamber 303, and a sample outlet 304; the microcavity sample inlet 301 is connected with the microcavity detection layer sample inlet connection hole 105, and the sample outlet hole 304 is connected with the microcavity detection layer sample outlet connection hole 106; the micropore reaction chamber 303 is connected with a microcavity sample inlet 301 and a sample outlet 304 through a flow channel 302.
The eluent liquid storage cavity (115) is provided with an eluent sample inlet hole (103).
As shown in fig. 5, this embodiment further provides a method for using a multilayer microfluidic chip integrating nucleic acid extraction, purification and digital detection, including:
(1) A detection preparation stage: first, 6 μ l of an aqueous elution reagent 125 (water phase) is injected into an eluent liquid storage cavity 115 through an eluent sample inlet 103, and then mineral oil 124 (oil phase) is injected into a lysate liquid storage cavity 111, and the mineral oil flows through a first washing liquid storage cavity 112, a second washing liquid storage cavity 113 and a mineral oil liquid storage cavity 114 through a bottom connecting pipeline 101 by tilting the chip and is filled. A sufficient amount of mineral oil should be maintained in each reservoir. The reagents used for extracting and purifying nucleic acid are nucleic acid lysate 121, first washing solution 122 and second washing solution 123 in sequence, and the reagents are injected into the lysate liquid storage cavity 111, the first washing solution liquid storage cavity 112 and the second washing solution liquid storage cavity 113 in sequence, wherein the volumes of the injected reagents are 8 mul, 4 mul and 4 mul respectively.
(2) Sampling: mu.l of sample was injected into the lysate reservoir 111.
(3) Nucleic acid extraction and purification: the nucleic acid lysis solution 121, the first washing solution 122 and the second washing solution 123 form reagent droplets in mineral oil 124, and the elution reagent 125 and the mineral oil 124 maintain a stable water-oil interface through the micro-column array 102 and are stably stored in the eluent storage cavity 115 through the capillary burst valve 104. Under the action of a magnet, the nucleic acid extraction magnetic beads 131 move longitudinally to mix with the lysis solution reagent 121, and move transversely to transfer nucleic acid molecules to the first washing solution 122 and the second washing solution 12 in sequence
3. Mineral oil 124, elution reagent 125. The nucleic acid molecules are released from the mixed solution of the lysis solution reagent 121 and the sample and combined with the nucleic acid extraction magnetic beads 131, residual impurities are removed by washing in the first washing solution 122 and the second washing solution 123, the nucleic acid molecules carried by the magnetic beads in the elution reagent 125 are eluted, and the preparation of the PCR reaction solution is completed.
(4) Micro-cavity sample introduction and oil sealing: after the elution process is finished, other liquid storage cavities except the mineral oil liquid storage cavity 114 are sealed by using an adhesive tape, the oil phase is continuously injected into the mineral oil liquid storage cavity 114 to exert pressure, the oil phase pushes the water phase to orderly enter the lower micro-cavity sample inlet 301 through the micro-cavity detection layer sample inlet connecting hole 105, the steps of filling and oil sealing of the reagent in the micro-pore reaction chamber 303 are finished through the flow channel 302, and the redundant reagent is discharged out of the chip through the sample outlet 304 and the micro-cavity detection layer sample outlet connecting hole 106.
(5) And (3) amplification detection: the nucleic acid extraction, purification and digital detection chip is placed in an in-situ PCR instrument and an amplification program is operated, and when the operation of the amplification program is finished and the device is cooled to room temperature, the chip is placed under a fluorescence microscope to observe the amplification result.
(6) And (3) data analysis: the fluorescence shot Image (figure 6) is analyzed by using Image, the initial concentration of the target nucleic acid in the sample to be detected can be obtained by counting the number of the positive micropores in each chip and combining the Poisson distribution principle, and thus, the qualitative and quantitative detection of the pathogen is realized.

Claims (6)

1. A multilayer micro-fluidic chip integrating nucleic acid extraction, purification and digital detection is characterized in that: the multilayer micro-fluidic chip comprises a PDMS-based nucleic acid extraction and purification layer (100), a glass substrate (200) and a silicon-based micro-cavity digital PCR detection layer (300) from top to bottom; the PDMS-based nucleic acid extraction and purification layer (100) consists of a liquid storage cavity (110), a bottom connecting pipeline (101), a micro-column array (102), a capillary explosion valve (104), a micro-cavity detection layer sample inlet connecting hole (105) and a micro-cavity detection layer sample outlet connecting hole (106); the liquid storage cavity (110) comprises a lysis solution storage cavity (111), a first washing solution storage cavity (112), a second washing solution storage cavity (113), a mineral oil storage cavity (114) and an eluent liquid storage cavity (115) which are sequentially connected through a bottom connecting pipeline (101); the micro-column array (102) is used for separating water and oil in a mineral oil storage cavity (114) and an eluent storage cavity (115); the capillary explosion valve (104) is used for preventing liquid in the eluent liquid storage cavity (115) from spontaneously entering the micro-cavity detection layer sample inlet connecting hole (105) in the nucleic acid purification process.
2. The multilayer microfluidic chip of claim 1, wherein: the lysate liquid storage cavity (111), the first washing liquid storage cavity (112) and the second washing liquid storage cavity (113) realize that the lysate, the first washing liquid and the second washing liquid are not interfered with each other by utilizing a water-in-oil principle.
3. The multi-layer microfluidic chip of claim 1, wherein: the glass substrate (200) is provided with holes corresponding to the sample inlet connecting hole (105) and the sample outlet connecting hole (106) of the microcavity detection layer.
4. The multilayer microfluidic chip of claim 1, wherein: the silicon-based microcavity digital PCR detection layer (300) comprises a microcavity sample inlet (301), a runner (302), a micropore reaction chamber (303) and a sample outlet (304); the microcavity sample inlet (301) is connected with the microcavity detection layer sample inlet connecting hole (105), and the sample outlet hole (304) is connected with the microcavity detection layer sample outlet connecting hole (106); the micropore reaction chamber (303) is connected with the microcavity sample inlet (301) and the sample outlet (304) through a flow channel (302).
5. The multi-layer microfluidic chip of claim 1, wherein: the eluent liquid storage cavity (115) is provided with an eluent sample inlet hole (103).
6. The use method of the multilayer microfluidic chip integrating nucleic acid extraction, purification and digital detection as claimed in claim 1 comprises the following steps:
(1) Firstly, an aqueous elution reagent is pre-filled into an eluent liquid storage cavity (115), then mineral oil is injected into a lysate liquid storage cavity (111), and the mineral oil flows through a first washing liquid storage cavity (112), a second washing liquid storage cavity (113) and a mineral oil liquid storage cavity (114) through a bottom connecting pipeline (101) and is filled through an inclined chip; injecting nucleic acid extracting and purifying reagents into corresponding liquid storage cavities, wherein each reagent is separated in an independent cavity;
(2) In the detection process, a sample is added into the lysate liquid storage cavity (111), cells or viruses in the sample are cracked, released nucleic acid molecules are adsorbed by the nucleic acid extraction magnetic beads (131), and then the nucleic acid extraction magnetic beads (131) are dragged into a subsequent reaction chamber to finish washing and elution processes, so that a PCR reaction liquid is obtained;
(3) The adhesive tape seals other liquid storage cavities except the mineral oil liquid storage cavity (114), and the oil phase continuously injects oil phase into the mineral oil liquid storage cavity (114) to exert pressure to push the water phase to enter the silicon-based micro-cavity digital PCR detection layer (300) orderly through the micro-cavity detection layer sample inlet connecting hole (105); finally, the chip is placed in an in-situ PCR instrument, and relevant programs are set to observe the detection result under a fluorescence microscope after the reaction is finished.
CN202211031355.8A 2022-08-26 2022-08-26 Multilayer micro-fluidic chip integrating nucleic acid extraction, purification and digital detection Pending CN115491297A (en)

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CN202211031355.8A CN115491297A (en) 2022-08-26 2022-08-26 Multilayer micro-fluidic chip integrating nucleic acid extraction, purification and digital detection

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CN202211031355.8A CN115491297A (en) 2022-08-26 2022-08-26 Multilayer micro-fluidic chip integrating nucleic acid extraction, purification and digital detection

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