CN218435756U - Nucleic acid detection micro-fluidic chip for in-situ capture and amplification and nucleic acid detector - Google Patents

Abstract

The utility model provides a nucleic acid detection micro-fluidic chip and nucleic acid detector of normal position capture and amplification, wherein detect the chip, include: a shell, a cleaning liquid pipe, a waste liquid pool pipe and a reagent pipe which are respectively sealed are arranged in the shell, the outer side of the shell is provided with a sample tube which is preset with nucleic acid cracking liquid; the flow channel plate is connected to the bottom of the shell, a flow channel is formed in the flow channel plate, a reaction cavity is formed in the flow channel plate, and nucleic acid capture filter paper is arranged in the reaction cavity; the content in the sample tube can be controlled to enter the reaction chamber through the flow channel, and the nucleic acid capture is realized at the nucleic acid capture filter paper; the content in the washing liquid pipe can be controlled to enter the reaction chamber through the flow channel to wash the captured nucleic acid; the content in the reagent tube can be controlled to enter the reaction chamber through the flow channel; the contents of the reaction chamber can be controlled to be stored in the waste liquid pool tube. The utility model discloses a nucleic acid sample pretreatment, nucleic acid schizolysis, nucleic acid capture, nucleic acid washing, nucleic acid amplification's design of integrating.

Description

Nucleic acid detection micro-fluidic chip for in-situ capture and amplification and nucleic acid detector

Technical Field

The utility model belongs to the technical field of biological analysis detects chip design, concretely relates to nucleic acid detection micro-fluidic chip and nucleic acid detector of normal position capture and amplification.

Background

The traditional nucleic acid detection methods such as Southern blot hybridization, agarose gel electrophoresis and the like have the defects of time and labor waste, low sensitivity, difficulty in realizing high-throughput detection, even need of using radioactive reagents and the like. Nucleic acid amplification methods such as polymerase chain reaction PCR, loop-mediated isothermal amplification LAMP, recombinase polymerase amplification RPA and the like can greatly improve the detection sensitivity, and simultaneously become common methods for modern molecular biology experiments due to relatively simple operation. Pathogen detection by nucleic acid amplification generally comprises four steps of sample pretreatment, nucleic acid extraction, amplification and detection. The traditional method for detecting pathogens in a laboratory needs professionals to complete in the laboratory with professional conditions, needs manual operation to complete reagent transfer and addition, still needs to complete a large amount of complicated operations, possibly exposes reaction reagents in the air in the operation process, is extremely easy to cause pollution, and needs to improve accurate temperature control by means of a temperature controller, so that the traditional method for detecting nucleic acids in the laboratory is limited to realize quick and sensitive detection of pathogens.

The microfluidic chip technology (Microfluidics) integrates basic operation units of sample preparation, reaction, separation, detection and the like in biological, chemical and medical analysis processes into a micro-scale chip to automatically complete the whole analysis process. Due to its great potential in the fields of biology, chemistry, medicine, etc., it has been developed as one of the important vector tools for nucleic acid detection. The microfluidic chip has many references on nucleic acid extraction amplification detection, but most of the microfluidic chips still stay in the laboratory stage. The reason is that various micro valves, micro pumps and auxiliary equipment are needed, the complexity of chip application is increased, the manufacturing difficulty and cost are greatly increased, the further clinical application of the chip is limited, and the fully integrated pathogen nucleic acid detection is difficult to realize, so that a fully integrated and high-sensitivity micro-fluidic chip is urgently needed, the nucleic acid detection process is improved, and the detection time and the chip manufacturing difficulty and cost are reduced.

SUMMERY OF THE UTILITY MODEL

Therefore, the to-be-solved technical problem of the present invention is to provide a nucleic acid detection micro-fluidic chip and a nucleic acid detector for in-situ capture and amplification, which can realize the fully integrated and highly sensitive detection of nucleic acid of pathogenic microorganisms.

In order to solve the above problems, the present invention provides a nucleic acid detection micro-fluidic chip for in-situ capture and amplification, comprising:

a shell, wherein a cleaning liquid pipe, a waste liquid pool pipe and a reagent pipe which are respectively sealed are arranged in the shell, a sample tube for presetting nucleic acid lysate is arranged on the outer side of the shell;

the flow channel plate is connected to the bottom of the shell, a flow channel is constructed in the flow channel plate, a reaction cavity is further constructed on the flow channel plate, and nucleic acid capture filter paper is arranged in the reaction cavity;

the content in the sample tube can be controlled to enter the reaction chamber through the flow channel, and the nucleic acid capture is realized at the nucleic acid capture filter paper, and/or a pressing ring is further arranged in the reaction chamber, the pressing ring is used for fixing the nucleic acid capture filter paper, and the pressing ring is of a C-ring structure;

the content in the washing liquid pipe can be controlled to enter the reaction chamber through the flow channel to wash the captured nucleic acid;

the content in the reagent tube can be controlled to enter the reaction chamber through the flow channel;

the content in the reaction chamber can be controlled and stored in the waste liquid pool pipe.

In some embodiments, still be provided with many hollow thimbles on the runner board, the runner with position department that wash liquor pipe, waste liquid pond pipe, reagent pipe respectively correspond respectively is equipped with one hollow thimble, just the bottom of wash liquor pipe, reagent pipe, waste liquid pond pipe all has lower sealed plug, lower sealed plug has the parcel and seals the shutoff position and the hollow thimble of hollow thimble wear out so that corresponding pipe valve with the intercommunication position of runner or cavity intercommunication.

In some embodiments, the cleaning liquid pipe, the reagent pipe and the waste liquid pool pipe are all provided with an upper sealing rubber plug, and the upper sealing rubber plug is arranged in the upper area of the lower sealing rubber plug and can be controlled to move close to the lower sealing rubber plug.

In some embodiments, a hard layer is disposed on a side of the upper sealing rubber plug facing away from the lower sealing rubber plug.

In some embodiments, the flow field plate has a protruding portion protruding from an outer sidewall of the housing, and the reaction chamber is configured on the protruding portion, and the protruding portion has a first side and a second side opposite to each other, wherein the first side corresponds to the temperature control member, and the second side corresponds to the fluorescence detection member.

In some embodiments, the first side has a recess thereon, and the reaction chamber is in a region corresponding to the recess.

In some embodiments, the outer edge of the protruding portion is provided with a chip catch.

In some embodiments, a tube rack is disposed within the housing, and the wash solution tube, the reagent tube, and the waste reservoir tube are disposed within the tube rack.

In some embodiments, the top opening of the housing is covered with a cover plate.

The utility model also provides a nucleic acid detector, including the push rod, the push rod uses with the cooperation of the nucleic acid detection micro-fluidic chip of foretell normal position capture and amplification, works as when the nucleic acid detection micro-fluidic chip of normal position capture and amplification includes sealed plug, hollow thimble, the push rod can the application of force in go up sealed plug to make the content in the corresponding pipe valve flow out or make it is corresponding to go up sealed plug parcel shutoff hollow thimble.

The utility model provides a pair of nucleic acid detection micro-fluidic chip of normal position capture and amplification and have its nucleic acid detection appearance, through with the cleaning solution pipe, waste liquid pond pipe, the sample pipe, the reagent pipe is through the flow and the storage of the content in the corresponding lumen of passageway basis nucleic acid extraction process control in proper order, nucleic acid sample pretreatment has been realized, the nucleic acid schizolysis, nucleic acid capture, nucleic acid cleaning, the design of integrating of nucleic acid amplification, this does benefit to and detects pathogen nucleic acid more fast, simultaneously owing to adopt nucleic acid capture filter paper to carry out the normal position to nucleic acid and catch and amplify and have higher sensitivity.

Drawings

Fig. 1 is a schematic view (partially disassembled) of a disassembled structure of a nucleic acid detection microfluidic chip for in-situ capture and amplification according to an embodiment of the present invention;

fig. 2 is a schematic view (partially disassembled) of another disassembled structure of the in-situ capturing and amplifying nucleic acid detecting microfluidic chip according to the embodiment of the present invention;

FIG. 3 is a schematic view (cross section) of the internal structure of the waste liquid tank tube in FIG. 1;

FIG. 4 is a schematic view of a disassembled structure of the sample tube of FIG. 1;

fig. 5 is a schematic top view perspective view of the flow field plate of fig. 1;

fig. 6 is a schematic perspective view of the flow channel plate in fig. 2 from a bottom view.

The reference numerals are represented as:

1. a housing; 101. a pipe frame; 102. a cover plate; 12. cleaning a liquid pipe; 13. a waste liquid pool pipe; 131. an air outlet; 14. a reagent tube; 15. a sample tube; 151. an upper end cap; 152. a fixed cover; 153. a sample tube body; 154. an aluminum gland; 2. a runner plate; 21. a flow channel; 22. a reaction chamber; 23. nucleic acid capture filter paper; 24. a hollow thimble; 25. chip buckling; 300. a lower sealing rubber plug; 301. and a rubber plug is sealed.

Detailed Description

Referring to fig. 1 to 6 in combination, according to an embodiment of the present invention, there is provided an in-situ capturing and amplifying nucleic acid detection microfluidic chip, including: the device comprises a shell 1, wherein a cleaning liquid pipe 12, a waste liquid pool pipe 13 and a reagent pipe 14 which are respectively sealed are arranged in the shell 1, and a sample pipe 15 for presetting nucleic acid cracking liquid is arranged on the outer side of the shell 1; a flow channel plate 2 connected to the bottom of the housing 1, wherein a flow channel 21 is formed in the flow channel plate 2, a reaction chamber 22 is formed on the flow channel plate 2, and a nucleic acid capture filter paper 23 is arranged in the reaction chamber 22; the content in the sample tube 15 can be controlled to enter the reaction chamber 22 through the flow channel 21, and the capture of nucleic acid is realized at the nucleic acid capture filter paper 23; the content in the washing liquid tube 12 can be controlled to enter the reaction chamber 22 through the flow channel 21 to wash the captured nucleic acid; the contents of the reagent tube 14 can be controlled to enter the reaction chamber 22 through the flow channel 21; the content can be controlled to be saved in waste liquid pond pipe 13 among reaction chamber 22, need explain that, through carrying out the amplification (adopt the control by temperature change part to realize the normal position amplification) and adopting corresponding fluorescence detection equipment (fluorescence detection part) to carry out corresponding detection to the sample after drawing in reaction chamber 22, can realize adopting the utility model discloses a chip is to the integrated purpose of the full flow of nucleic acid detection's sample preparation, nucleic acid extraction, nucleic acid amplification process.

In the technical scheme, the cleaning liquid pipe 12, the waste liquid pool pipe 13, the sample pipe 15 and the reagent pipe 14 sequentially control the outflow and storage of contents in corresponding pipe cavities according to a nucleic acid extraction process through the flow channel 21, so that the integrated design of nucleic acid sample pretreatment, nucleic acid cracking, nucleic acid capturing, nucleic acid cleaning and nucleic acid amplification is realized, the detection of pathogen nucleic acid is facilitated more quickly, and meanwhile, the nucleic acid in-situ capturing and amplification by adopting the nucleic acid capturing filter paper 23 have higher sensitivity.

Specifically, the sample tube 15 is used for collecting and lysing a sample to be tested (the sample may be DNA/RNA with a certain concentration, or a pathogenic microorganism sample including cells, viruses, tissues, bacteria, and fungi); the cleaning liquid tube 12 is used for cleaning the nucleic acid capturing filter paper 23 (arranged in the reaction chamber 22) after the nucleic acid is captured, and removing certain impurities and lysate residues; the reagent tube 14 is used for storing liquid reaction reagents, such as polymerase chain reaction PCR, recombinase polymerase reaction RPA, loop-mediated isothermal amplification LAMP, and nucleic acid sequence-dependent amplification NASBA reaction reagents; the reaction chamber 22 is used for performing nucleic acid amplification (in-situ amplification), such as Polymerase Chain Reaction (PCR), recombinase polymerase Reaction (RPA), loop-mediated isothermal amplification (LAMP), nucleic acid sequence-dependent amplification (NASBA), and the like, and fluorescent information of the amplified contents can be acquired by a fluorescent detection component matched with the reaction chamber; the waste liquid tank pipe 13 is used for storing waste liquid.

The nucleic acid capturing filter paper 23 is specifically a chitosan-modified capturing filter paper, which has hydrophilicity, and in some embodiments, a pressing ring (not shown in the figure) is further disposed in the reaction chamber 22, the pressing ring is used for fixing the nucleic acid capturing filter paper, and the pressing ring is a C-ring structure, so that when a reagent flows through the filter paper, the internal space of the pressing ring is filled first, and then the reagent flows to the side C-ring outlet, and it can be ensured that no bubble is present in the reaction chamber 22, and no interference is caused to the subsequent reaction.

In some embodiments, the flow channel plate 2 is further provided with a plurality of hollow thimbles 24, the plurality of hollow thimbles 24 are fixed through the needle seat holes, the positions of the flow channel 21 corresponding to the cleaning solution tube 12, the waste liquid pool tube 13 and the reagent tube 14 are respectively provided with one hollow thimble 24, the bottoms of the cleaning solution tube 12, the reagent tube 14 and the waste liquid pool tube 13 are all provided with a lower sealing rubber plug 300, and the lower sealing rubber plug 300 is provided with a blocking position for wrapping the sealing hollow thimbles 24 and a communication position for enabling the corresponding tube valve to communicate with the flow channel 21 or the cavity by penetrating the hollow thimbles 24. Specifically, an external pushing component (e.g., a push rod) pushes the relevant tube valve to move close to the hollow thimble 24 at the top of the lower sealing rubber plug 300, so as to switch the lower sealing rubber plug 300 from the plugging position to the communication position, that is, when the lower sealing rubber plug 300 is in the plugging position, the content in the corresponding tube valve is stored in the tube valve and cannot flow out, and when the lower sealing rubber plug 300 is in the communication position, the content in the corresponding tube valve is forced out to the reaction chamber 22 of the flow channel 21, so as to implement the corresponding nucleic acid extraction process, such as extraction, cleaning, reagent reconstitution, amplification reaction, and the like. In the technical scheme, the contents in the cavity valve can be discharged and blocked in a mode that the hollow thimble 24 pierces the lower sealing rubber plug 300, the process is consistent with the injection (namely pushing and pressing each cavity valve) action of the corresponding cavity valve, and the structure is simple and the control is simple and convenient. Furthermore, the chip adopts a thimble-piercing structure, and based on the piercing valve structure, various conventional reagents for nucleic acid detection can be integrated on a tiny chip unit, so that a brand-new nucleic acid detection mode is realized, and the requirements of long-term storage and long-distance transportation of the reagents can be met.

In some embodiments, the cleaning solution tube 12, the reagent tube 14, and the waste liquid pool tube 13 are all provided with an upper sealing rubber plug 301, and the upper sealing rubber plug 301 is located at an upper region of the lower sealing rubber plug 300 and can be controlled to move close to the lower sealing rubber plug 300, so that when the corresponding tube valve moves close to the hollow thimble 24 all the time, when the content in the corresponding tube valve completely flows out, the corresponding tube valve can be plugged, and the liquid in other processing procedures can be prevented from flowing back into the tube valve. In a preferred embodiment, one side of the upper sealing rubber plug 301 departing from the lower sealing rubber plug 300 is provided with a hard layer, for example, hard plastic is arranged on the top surface of the upper sealing rubber plug 301, so that the upper sealing rubber plug 301 forms a soft and hard combined rubber plug, specifically, the tail end of the external push rod can directly apply force on the top surface of the upper sealing rubber plug 301, the hard layer is arranged to effectively prevent the rubber plug from being unevenly deformed and not inclined in the force application process, and the corresponding pipe valve is guaranteed to smoothly, reliably and stably move downwards.

Specifically, referring to fig. 4, a decomposition structure of the sample tube 15 in an embodiment is specifically shown, which includes a sample tube body 153, where the sample tube body 153 is a tube structure penetrating from top to bottom, an upper sealing rubber plug 301 and a lower sealing rubber plug 300 are respectively plugged at upper and lower ends of the sample tube body 153, an interval is provided between the two rubber plugs in an initial state, the interval forms an accommodating space of the sample tube, an aluminum pressing cover 154 is further included outside the lower sealing rubber plug 300, and is matched with the lower sealing rubber plug 300 to form reliable sealing on a top end of the sample tube 15, a fixing cover 152 and an upper end cap 151 combined with the fixing cover 152 are further sleeved outside the upper sealing rubber plug 301, and the fixing cover 152 has an internal thread, which is combined with the upper end cap 151 to seal the sample tube 15.

Referring specifically to fig. 3, an internal structure of the waste liquid pool pipe 13 in an embodiment is shown, wherein a lower sealing rubber plug 300 and an upper sealing rubber plug 301 therein are arranged at an upper and lower interval, an interval between the two forms a liquid inlet channel of the waste liquid pool pipe 13, the gap is reduced until the upper sealing rubber plug 301 completely wraps and blocks a hollow thimble 24 below the upper sealing rubber plug in the downward moving process of the waste liquid pool pipe 13, so that the waste liquid pool pipe 13 is sealed, the waste liquid pool pipe 13 is separated from a reaction chamber 22, nucleic acid pollution caused by leakage of amplification products is avoided, and an air outlet 131 is formed in the top of the waste liquid pool pipe to ensure that each chamber can smoothly enter and exit liquid.

Referring to fig. 5 and 6, the flow field plate 2 has a protruding portion protruding from the outer wall of the housing 1, and the reaction chamber 22 is configured on the protruding portion, and the protruding portion has a first side surface and a second side surface opposite to each other, wherein the first side surface corresponds to the temperature control unit, and the second side surface corresponds to the fluorescence detection unit, that is, during the detection of nucleic acid amplification, the temperature control unit is disposed at the first side surface, and the fluorescence detection unit can be disposed at the second side surface, so that the arrangement of the fluorescence detection unit does not occupy the temperature control area (heated region) of the temperature control unit, thereby enabling the contents in the reaction chamber 22 to realize PCR cycling amplification. In another embodiment, the first side surface has a recess, and the reaction chamber 22 is located in the region corresponding to the recess, so as to reduce the heating wall thickness of the reaction chamber 22 when the reaction chamber is heated by the supporting instrument during the amplification reaction, thereby making the temperature of the reaction chamber during the nucleic acid amplification reaction more accurate.

In some embodiments, the outer edge of the protruding portion is provided with a chip clip 25, which can form a structure similar to a card slot of an "SD" card with a mating detection instrument, so that the chip can be fixed when the instrument is matched with the chip for sample processing and detection.

In some embodiments, a pipe rack 101 is disposed in the housing 1, the cleaning liquid pipe 12, the reagent pipe 14, and the waste liquid pool pipe 13 are disposed in the pipe rack 101, that is, the pipe rack 101 disposed in the housing 1 is disposed between an inner wall of the housing 1 and an outer wall of each pipe valve, and can effectively and reliably fix each pipe valve, at this time, the housing 1 has an effect of improving the aesthetic appearance of the whole chip, the top opening of the housing 1 is covered and connected with a cover plate 102, and each pipe valve in the housing 1 can be prevented from coming off from the top opening, thereby preventing loss of each pipe valve inside, and ensuring reliable and stable positions of each pipe valve inside.

The utility model also provides a nucleic acid detector, including the push rod, the push rod uses with the nucleic acid detection micro-fluidic chip cooperation of foretell normal position capture and amplification, and when the nucleic acid detection micro-fluidic chip of normal position capture and amplification included last sealed plug 301, hollow thimble 24, the push rod can apply force in last sealed plug 301 to make the content in the corresponding pipe valve flow out or make and go up sealed plug 301 parcel corresponding hollow thimble 24 of shutoff.

The technical solution of the present invention is further explained with reference to a specific embodiment.

Taking the detection of the oral swab new coronavirus by PCR as an example, the specific implementation mode of the chip is as follows:

storing a reagent: each independent reagent tube can be pre-stored with an upper rubber plug (namely, the upper sealing rubber plug 301, the same below) and a lower rubber plug (namely, the lower sealing rubber plug 300, the same below) for subsequent experiments, the reagent in the cleaning solution chamber (namely, the cleaning solution tube 12, the same below) can be 1ml EPC water (DNase, RNase free) (product number: R0022, manufacturer: byunyun), and the storage reagent in the reagent tube (namely, the reagent tube 14, the same below) can be MIX reagent for PCR amplification;

collecting samples: collecting a sample by using a smear swab, directly rinsing the collected sample in the sample tube 15, and releasing nucleic acid in the sample tube 15 after taking out the swab; in this experiment, a novel coronavirus (2019-nCoV) pseudovirus ribonucleic acid standard substance was selected instead (Cat number: NIM-RM5203, manufacturer: chinese institute of metrology science).

Nucleic acid release: the storage solution in the sample tube 15 is a lysis solution, which, once the sample has been introduced, can be lysed by a pathogen, releasing nucleic acids.

And (3) nucleic acid capture: the instrument is matched (the instrument structure is not unique, and only the following three conditions are met, namely, a push rod with motor control can form a structure similar to an injector with a rubber plug (namely the upper sealing rubber plug 301) in a liquid tube of a detection chip for liquid control, a temperature control module (namely a temperature control part) can be used for temperature required by reaction, a fluorescence detection module (namely a fluorescence detection part) is used for reading fluorescence at a reaction cavity 22 of the detection chip and nucleic acid capture filter paper 23 in real time, the push rod pushes the soft and hard combined rubber plug in the sample tube 15 to carry out plunger type pressure driving, the rubber plug in the tube is pushed downwards to inject, the sample tube is firstly moved downwards for a certain distance of travel, a thimble (namely a hollow thimble 24 and the lower thimble) of the sample tube fixed on a lower pipeline layer (namely the flow channel plate 2 and the lower pipeline) penetrates through the rubber plug (namely the lower sealing rubber plug 300 and the lower pipeline) at the lower end of the sample tube, fluid can enter a fluid pipeline layer of the lower pipeline layer, the fluid tube further flows through the nucleic acid capture plate 23 to carry out nucleic acid capture, the fluid can enter a waste liquid pool (namely the waste liquid pool and the filter paper and the cleaning fluid in the liquid pool, and the filter paper tube 13 is closed.

Nucleic acid washing: the cooperation instrument push rod promotes the rubber stopper of the soft or hard combination in the washing liquid pipe and carries out plunger formula pressure drive, during the injection, can let the stroke that washing liquid pipe moved certain distance down earlier, make fixed washing liquid thimble on the lower pipeline layer puncture washing liquid pipe lower extreme plug, and then can let washing liquid flow in the fluid pipeline, the impurity cleaning on the filter paper is carried out to the filter paper that flows again, wash the impurity on the filter paper, can enter into the waste liquid pond after liquid flows through filter paper, this in-process sample tube is the closed condition with the reagent pipe.

Nucleic acid amplification: the matching instrument push rod pushes the soft and hard combined rubber plug in the reagent tube to perform plunger type pressure driving, fluid in the reagent tube is squeezed into the reaction chamber, then the matching instrument push rod pushes the waste liquid pool down for a distance, the waste liquid pool switch valve component (namely the lower sealing rubber plug is in a state of wrapping and blocking the hollow thimble 24) is closed, and then the matching instrument push rod pushes the rubber plug in the reagent tube at the bottom end of the tube down, so that the sealing at two ends of the reaction chamber can be realized. Then the nucleic acid amplification can be carried out by matching with an instrument to carry out temperature-changing temperature control. When the reagent tube is driven to flow, the sample tube and the cleaning liquid tube are both in a closed state.

Outputting a detection result: and (3) in cooperation with a fluorescence detection module of a fully integrated instrument, carrying out real-time fluorescence detection on a reaction chamber and nucleic acid capture filter paper in a pipeline layer of a lower part of the detection chip, continuously drawing a fluorescence curve, and finally carrying out quantitative judgment according to a Ct value.

It is readily understood by a person skilled in the art that the advantageous ways described above can be freely combined, superimposed without conflict.

The above description is only exemplary of the present invention and should not be construed as limiting the present invention, and any modifications, equivalents and improvements made within the spirit and principles of the present invention are intended to be included within the scope of the present invention. The above is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, a plurality of modifications and variations can be made without departing from the technical principle of the present invention, and these modifications and variations should also be regarded as the protection scope of the present invention.

Claims (10)

1. The nucleic acid detection micro-fluidic chip for in-situ capture and amplification is characterized by comprising:

the device comprises a shell (1), wherein a cleaning liquid pipe (12), a waste liquid pool pipe (13) and a reagent pipe (14) which are sealed respectively are arranged in the shell (1), and a sample pipe (15) for presetting nucleic acid cracking liquid is arranged on the outer side of the shell (1);

a flow channel plate (2) connected to the bottom of the housing (1) and having a flow channel (21) formed therein, wherein a reaction chamber (22) is further formed on the flow channel plate (2), and a nucleic acid capture filter paper (23) is formed in the reaction chamber (22);

the content in the sample tube (15) can be controlled to enter the reaction chamber (22) through the flow channel (21) and capture nucleic acid at the nucleic acid capture filter paper (23); and/or a pressing ring is also arranged in the reaction chamber (22), the pressing ring is used for fixing the nucleic acid capture filter paper (23), and the pressing ring is of a C-ring structure;

the content in the washing liquid pipe (12) can be controlled to enter the reaction chamber (22) through the flow channel (21) to wash the captured nucleic acid;

the content of the reagent tube (14) can be controlled to enter the reaction chamber (22) through the flow channel (21);

the contents of the reaction chamber (22) can be controlled to be stored in the waste reservoir tube (13).

2. The microfluidic chip for nucleic acid detection according to claim 1, wherein the flow channel plate (2) is further provided with a plurality of hollow ejector pins (24), the positions of the flow channel (21) corresponding to the cleaning liquid tube (12), the waste liquid pool tube (13) and the reagent tube (14) are respectively provided with one hollow ejector pin (24), the bottoms of the cleaning liquid tube (12), the reagent tube (14) and the waste liquid pool tube (13) are respectively provided with a lower sealing rubber plug (300), and the lower sealing rubber plug (300) has a plugging position for wrapping and sealing the hollow ejector pins (24) and a communication position for allowing the corresponding tube valve to communicate with the flow channel (21) or the chamber by penetrating the hollow ejector pins (24).

3. The microfluidic chip for nucleic acid detection according to claim 2, wherein the upper sealing rubber plug (301) is disposed in the upper region of the lower sealing rubber plug (300) and can be controlled to move close to the lower sealing rubber plug (300) in the wash liquid tube (12), the reagent tube (14) and the waste liquid pool tube (13).

4. The microfluidic chip for nucleic acid detection according to claim 3, wherein a hard layer is disposed on a side of the upper sealing rubber plug (301) facing away from the lower sealing rubber plug (300).

5. The microfluidic chip for nucleic acid detection according to claim 1, wherein the flow field plate (2) has a protruding portion protruding from an outer sidewall of the housing (1), and the reaction chamber (22) is configured on the protruding portion, and the protruding portion has a first side and a second side opposite to each other, wherein the first side corresponds to the temperature control part and the second side corresponds to the fluorescence detection part.

6. The nucleic acid detecting microfluidic chip according to claim 5, wherein the first side surface has a recess, and the reaction chamber (22) is located in a region corresponding to the recess.

7. The microfluidic chip for nucleic acid detection by in situ capture and amplification according to claim 5, wherein the outer edge of the protruding portion is provided with a chip latch (25).

8. The microfluidic chip for nucleic acid detection according to claim 1, wherein a tube rack (101) is disposed in the housing (1), and the wash solution tube (12), the reagent tube (14), and the waste solution pool tube (13) are disposed in the tube rack (101).

9. The nucleic acid detection microfluidic chip according to claim 8, wherein the top opening of the housing (1) is covered with a cover plate (102).

10. A nucleic acid detecting instrument, comprising a push rod, wherein the push rod is used in cooperation with the nucleic acid detecting microfluidic chip of any one of claims 1 to 9, and when the in-situ capturing and amplifying nucleic acid detecting microfluidic chip comprises an upper sealing rubber plug (301) and a hollow thimble (24), the push rod can apply force to the upper sealing rubber plug (301) to enable the contents in the corresponding tube valve to flow out or enable the upper sealing rubber plug (301) to wrap and seal the corresponding hollow thimble (24).

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