CN114350511A

CN114350511A - Cassette and method for rapidly detecting nucleic acid

Info

Publication number: CN114350511A
Application number: CN202111654510.7A
Authority: CN
Inventors: 黄超杰
Original assignee: Guangdong Runpon Bioscience Co Ltd
Current assignee: Guangdong Runpon Bioscience Co Ltd
Priority date: 2021-12-30
Filing date: 2021-12-30
Publication date: 2022-04-15

Abstract

The invention discloses a nucleic acid rapid detection card box and a method, the nucleic acid rapid detection card box comprises a reagent plate, a shell, a magnetic rod sleeve, a plurality of piston rods and a plurality of control valves, the reagent plate is movably connected with the shell, an extraction chamber used for extracting nucleic acid is formed between the reagent plate and the shell, the top of the reagent plate is concavely provided with a plurality of piston channels and a plurality of storage chambers, the plurality of storage chambers are annularly arranged at the periphery of the position of the piston channel, the shell is provided with a magnetic rod hole, a sample adding hole, an air vent and a piston hole corresponding to the plurality of piston channels, the magnetic rod hole, the sample adding hole and the air vent are arranged at the periphery of the piston hole and are used for communicating the inside and the outside of the extraction chamber, the magnetic rod sleeve is rotatably arranged on the magnetic rod hole, one end of the piston rod is positioned outside the piston hole, the other end of the piston rod passes through the piston hole and is slidably arranged in the piston channel, the piston channel is communicated with at least one storage chamber, the piston channel is provided with a space used for amplification and detection, the control valve is arranged on the reagent plate and used for controlling the on-off between the piston channel and the storage cabin.

Description

Cassette and method for rapidly detecting nucleic acid

Technical Field

The invention relates to the technical field of nucleic acid detection, in particular to a nucleic acid rapid detection card box and a method.

Background

Since the outbreak of the new coronavirus epidemic situation in 2019, the global demand of all countries for molecular detection technologies including the fluorescence quantitative qPCR technology shows the increase of the well-spraying type, and the traditional molecular detection steps mainly comprise the steps of nucleic acid extraction, reagent subpackaging, qPCR on-machine testing, result analysis and the like.

However, these procedures require the above-mentioned detection procedures to be performed in a professional physically partitioned qPCR molecular laboratory, and then the laboratory needs to be equipped with expensive instruments including nucleic acid extractors, biosafety cabinets, fluorescence quantitative qPCR instruments, etc., and then needs to rely on the operation of specially trained professional operators to perform the detection. Generally speaking, the traditional molecular detection reagent has the problems that the detection platform is expensive to build and operate, the equipment is expensive, professional operators and manual operation are complicated, the detection period is long, pollution is easy to occur, the risk of harm to the safety of the operators is generated, and the like, so that only part of three hospitals or large hospitals are qualified, qualified and capable of developing related molecular detection projects, many two hospitals and basic medical institutions cannot develop related detection projects, and after a new crown epidemic situation outbreak, the detection and the field detection of the basic hospitals are the key points for preventing the spread of the epidemic situation of the infectious diseases.

Disclosure of Invention

The invention aims to provide a card box and a method for quickly detecting nucleic acid, which aim to solve the technical problems that the prior nucleic acid molecule detection technology is qualified, qualified and capable only in part of three hospitals or large hospitals to develop related molecule detection items, and a plurality of hospitals and basic medical institutions cannot develop related detection items, so that the detection cost is high and the detection operation is difficult.

The invention provides a nucleic acid rapid detection card box, which comprises a reagent plate, a shell, a magnetic rod sleeve, a plurality of piston rods and a plurality of control valves, wherein the reagent plate is movably connected with the shell, an extraction chamber for extracting nucleic acid is formed between the reagent plate and the shell, a plurality of piston channels and a plurality of storage chambers are concavely arranged at the top of the reagent plate in the extraction chamber, the plurality of storage chambers are annularly arranged at the peripheries of the positions of the plurality of piston channels, a magnetic rod hole, a sample adding hole, an air vent and a piston hole corresponding to the positions of the plurality of piston channels are arranged at the top of the shell corresponding to the reagent plate, the magnetic rod hole, the sample adding hole and the air vent are arranged at the peripheries of the piston holes and are used for communicating the interior and the exterior of the extraction chamber, the magnetic rod sleeve is rotatably arranged on the magnetic rod hole and is arranged in the extraction chamber, the one end of piston rod is located outside the piston hole, the other end pass behind the piston hole slidable mounting in the piston passageway, the shell can for the reagent board rotates in order to drive the bar magnet hole respectively correspond to two at least different storage cabins in a plurality of storage cabins, the shell can for the reagent board removes in order to change at least one in bar magnet hole, the hole of adding the appearance and the bleeder vent with the distance between the top of reagent board, piston passageway and at least one the storage cabin intercommunication, piston passageway has the space that is used for augmenting and detecting, the control valve install in on the reagent board and be used for controlling the piston passageway with the break-make of intercommunication state between the storage cabin.

Further, the reagent plate is convexly provided with an amplification plate, an amplification pipeline is arranged in the amplification plate, the piston channel comprises the amplification pipeline, and the amplification pipeline is provided with a space for amplification and detection.

Further, the reagent board still have with the base that the shell is connected, the amplification board sets up on the base, piston channel's quantity is one, the middle part top-down of base is equipped with guiding hole and mounting hole in proper order, the storage cabin is concave to be located on the base, the amplification board install in on the mounting hole, still be equipped with on the base with the microchannel of storage cabin intercommunication, the one end of amplification pipeline with guiding hole intercommunication, the other end with the microchannel intercommunication, guiding hole, amplification pipeline and microchannel constitute piston channel, the control valve install in on the base and be used for controlling the break-make of microchannel.

Furthermore, the amplification pipeline comprises a guide channel, a first liquid guide pipe, an amplification chamber and a second liquid guide pipe which are sequentially communicated, the amplification chamber is used for providing the space for amplification and detection, the guide channel is communicated with the guide hole, and the second liquid guide pipe is communicated with the micro-channel.

Furthermore, a plurality of the storage cabins comprise a first chamber for storing the split binding solution, a second chamber for storing the first washing solution, a third chamber for storing the second washing solution, a fourth chamber for storing the eluent, a fifth chamber for storing the magnetic rod sleeve and a sixth chamber for storing the magnetic beads, wherein the first chamber, the second chamber, the third chamber, the fourth chamber, the fifth chamber and the sixth chamber are arranged in an isolated mode, and the fourth chamber is communicated with the micro flow channel.

Furthermore, the number of the control valves is one, the micro flow channel comprises a first micro flow pipe communicated with the second liquid guide pipe and a second micro flow pipe communicated with the fourth cavity, the control valves are one-way valves, one end of each control valve is communicated with the first micro flow pipe, and the other end of each control valve is communicated with the second micro flow pipe.

Furthermore, the storage cabins comprise a first chamber for storing the lysis binding solution, a second chamber for storing the first washing solution, a third chamber for storing the second washing solution, a fourth chamber for storing the eluent, a fifth chamber for storing the magnetic rod sleeve, a sixth chamber for storing the magnetic beads and a seventh chamber for storing the amplification freeze-drying reagent, wherein the first chamber, the second chamber, the third chamber, the fifth chamber and the sixth chamber are arranged in an isolated manner, and the fourth chamber and the seventh chamber are communicated with the micro-channel.

Further, the number of the control valve is one, the micro flow channel comprises a third micro flow pipe communicated with the second liquid guide pipe, a fourth micro flow pipe communicated with the fourth chamber and a fifth micro flow pipe communicated with the seventh chamber, the control valve is a three-way valve, one end of the control valve is communicated with the third micro flow pipe, and the other end of the control valve is communicated with the fourth micro flow pipe and the fifth micro flow channel respectively.

Further, the number of the control valves is two, the micro flow channel comprises a third micro flow pipe communicated with the second liquid guide pipe, a fourth micro flow pipe communicated with the fourth cavity and a fifth micro flow pipe communicated with the seventh cavity, the fourth micro flow pipe and the fifth micro flow pipe are communicated with the third micro flow pipe, the control valves are all one-way valves, one is used for controlling the on-off of the fourth micro flow pipe, and the other is used for controlling the on-off of the fifth micro flow pipe.

Further, the piston rod includes the body of rod and first sealing washer, the body of rod insert in the piston passageway, the concave annular first mounting groove that is equipped with on the lateral wall of the body of rod, the inner of first sealing washer install in the first mounting groove, outer end butt in the inner wall of piston passageway.

Further, the bottom of shell is equipped with in a recess and holds the chamber, reagent board card insert in hold the intracavity, bar magnet hole, application of sample hole, bleeder vent and piston hole all with hold the chamber intercommunication.

Further, the top of reagent board is the concave annular chamber that is equipped with still, piston passageway is located respectively with the storage cabin the inside and outside both sides of annular chamber, the shell includes annular roof, certainly the outward flange of annular roof is buckled the annular bounding wall that extends and is located in the annular bounding wall and with the registration arm of the coaxial setting of annular bounding wall, piston hole, bar hole, application hole and bleeder vent are all located on the annular roof, the registration arm with the annular roof is connected, the internal diameter of registration arm is greater than the internal diameter of piston hole, bar hole, application hole and bleeder vent all are located between registration arm and the annular bounding wall, the registration arm insert in the annular chamber, the diapire of annular roof and the inner wall of annular bounding wall enclose to be synthetic hold the chamber.

Further, the quick nucleic acid detection card box also comprises a second sealing ring, an annular second mounting groove is concavely arranged on the outer side wall of the reagent plate, the second mounting groove is close to the annular top plate, and the inner end of the second sealing ring is arranged in the second mounting groove, the outer end of the second sealing ring is in sliding fit with the annular enclosing plate.

Further, the quick nucleic acid detection card box also comprises a third sealing ring, an annular third mounting groove is concavely arranged on the outer side wall of the positioning tube, the third mounting groove is far away from the annular top plate, and the inner end of the third sealing ring is arranged in the third mounting groove, the outer end of the third sealing ring is in sliding fit with the inner wall of the annular cavity.

Further, the bar magnet cover includes that the magnetism inhales pole, spacing collar plate and puncture portion, the protruding bottom of inhaling the pole of puncture portion located the magnetism, the protruding lateral wall of inhaling the pole of spacing collar plate is located on the magnetism, the spacing collar plate rotate install in on the bar magnet hole, the magnetism has of inhaling the pole the one end of puncture portion insert in the storage compartment.

Further, the bar magnet cover still includes prevents losing the circle, prevent losing the circle and be cyclic annular, prevent losing the protruding bottom of locating the pole is inhaled to the magnetism, prevent losing the internal diameter of circle and follow and keep away from the one end of pole is inhaled towards being close to the magnetism the one end of pole reduces gradually, the puncture position is located prevent losing in the circle, prevent losing keeping away from of circle the one end of pole is inhaled to the magnetism extremely the axial distance of pole is inhaled to the magnetism is less than keep away from of puncture portion the one end of pole is inhaled to the magnetism extremely the axial distance of pole is inhaled to the magnetism.

Further, the magnetic rod sleeve further comprises a plurality of fin wings, the fin wings are convexly arranged on the outer side wall of the magnetic suction rod, and the fin wings are located between the limiting ring plate and the anti-lost ring and are close to the anti-lost ring.

The invention also provides a method for quickly detecting nucleic acid, which comprises the above-mentioned card box for quickly detecting nucleic acid, wherein each reagent is respectively stored in each storage cabin and each amplification cabin, and each storage cabin and each amplification cabin are in a sealed state, the control valve is in a closed state, and the method comprises the following steps:

step a, adding a proteinase K solution and a sample to be detected into a first chamber through a sample adding hole, and sealing the sample adding hole, wherein the sample to be detected has nucleic acid;

b, lifting the reagent plate to form a sealed extraction chamber, rotating the reagent plate or the shell, inserting the magnetic rod sleeve into the first chamber for stirring, and heating the sample to be detected in the first chamber to crack the sample to be detected so as to release nucleic acid;

c, adsorbing the magnetic beads in the sixth chamber into the first chamber by using a magnetic rod sleeve through the extraction chamber, wherein the reagent in the sixth chamber contains the magnetic beads, and adsorbing nucleic acid by using the magnetic beads;

d, adsorbing the magnetic beads with the nucleic acids in the first chamber into a reagent in a second chamber through an extraction chamber, wherein the reagent in the second chamber can remove foreign proteins on the magnetic beads;

e, adsorbing the magnetic beads with the nucleic acids in the second chamber into a reagent in a third chamber through the extraction chamber, wherein the reagent in the third chamber can remove salt ions on the magnetic beads;

f, adsorbing the magnetic beads with the nucleic acids in the third chamber into a reagent in a fourth chamber through an extraction chamber, heating the liquid in the fourth chamber, separating the nucleic acid molecules captured on the magnetic beads by the reagent in the fourth chamber, and removing the magnetic beads out of the fourth chamber by using a magnetic rod sleeve;

and step g, opening the control valve, pulling up the piston rod to adsorb nucleic acid molecules in the fourth chamber into the amplification chamber, closing the control valve, carrying out amplification reaction on the nucleic acid and reagents in the amplification chamber, and collecting fluorescence signals of the amplification chamber.

Further, the reagent is respectively stored in each storage cabin and each amplification cabin, and the specific steps that each storage cabin and each amplification cabin are in a sealed state are as follows:

the reagent board is dismantled, save the schizolysis bonding solution in first cavity and use the aluminium membrane heat-seal, save first washing liquid in the second cavity and use the aluminium membrane heat-seal, save the second washing liquid in the third cavity and use the aluminium membrane heat-seal, save the eluant in the fourth cavity and use the aluminium membrane heat-seal, save the magnetic bead solution in the sixth cavity and use the aluminium membrane heat-seal, save the freeze-drying reagent of amplification in the amplification warehouse and seal through piston rod and control valve, insert the reagent board in the shell.

Furthermore, a semi-permeable membrane which is permeable and impermeable to water is arranged on the air holes.

Compared with the prior art, the invention has the beneficial effects that:

the invention can add each reagent into each storage cabin in advance and seal, reduces the time of detection period, the air hole can be made very small, the air hole can be also provided with a permeable and impermeable semipermeable membrane, the sample hole is sealed at the later stage, the nucleic acid rapid detection card box can be put on a detection instrument with lifting and rotating functions, the magnetic rod sleeve is carried out in a sealed extraction chamber in the process of rising along with the shell and entering into another storage cabin, can prevent pollution, the detection instrument is operated in the whole process after inputting the program, reduces the complexity of manual operation, reduces the risk of harm of the nucleic acid to operators, the amplification pipeline of the piston channel can provide space for amplification and detection of qPCR amplification detection, the cost of the nucleic acid rapid detection card box is low, and related detection items can be developed in hospitals and basic medical institutions, the detection cost is low, the detection operation is simple, and the method is suitable for large-scale popularization.

Drawings

FIG. 1 is a first schematic view of a cassette for rapid nucleic acid detection according to an embodiment of the present invention;

FIG. 2 is a cross-sectional view taken along A-A in FIG. 1;

FIG. 3 is an enlarged view of a portion of FIG. 2 at B;

FIG. 4 is an exploded view of the nucleic acid rapid detection cartridge of FIG. 1;

FIG. 5 is a schematic structural diagram of a second embodiment of the kit for rapid nucleic acid detection according to the present invention;

FIG. 6 is an exploded view of the nucleic acid rapid detection cartridge of FIG. 5;

FIG. 7 is a third schematic view of a nucleic acid rapid detection cartridge according to an embodiment of the present invention;

FIG. 8 is a cross-sectional view taken along line C-C of FIG. 7;

FIG. 9 is a schematic structural diagram of a reagent plate according to a first embodiment of the present invention;

fig. 10 is a cross-sectional view taken along line D-D in fig. 9.

FIG. 11 is a schematic diagram of an amplification plate according to an embodiment of the present invention;

FIG. 12 is a cross-sectional view taken along line E-E of FIG. 11;

FIG. 13 is a schematic structural diagram of a control valve according to an embodiment of the present invention;

FIG. 14 is an exploded view of the control valve of FIG. 13;

fig. 15 is a schematic structural diagram of a magnetic rod sleeve according to an embodiment of the present invention;

FIG. 16 is a schematic structural diagram of a reagent plate according to a second embodiment of the present invention;

FIG. 17 is a cross-sectional view taken along line F-F of FIG. 16;

FIG. 18 is a schematic diagram of the amplification data of a mock sample for detecting influenza A virus pseudovirus according to the present invention.

In the figure:

10. a reagent plate; 11. a piston channel; 111. an amplification conduit; 1111. a guide channel; 1112. a first catheter; 1113. an amplification chamber; 1114. a second catheter; 12. a storage compartment; 121. a first chamber; 122. a second chamber; 123. a third chamber; 124. a fourth chamber; 125. a fifth chamber; 126. a sixth chamber; 127. a seventh chamber; 13. an amplification plate; 131. a columnar body; 132. a stationary ring plate; 133. a flat body; 14. a base; 141. a guide hole; 142. mounting holes; 143. a micro flow channel; 1431. a first microchannel; 1432. a second microchannel; 144. a valve through hole; 15. a fourth seal ring; 16. an annular cavity; 20. a housing; 21. an accommodating chamber; 22. a magnet bar hole; 23. a piston bore; 24. a sample application hole; 25. air holes are formed; 26. an annular top plate; 27. an annular coaming; 28. a positioning tube; 30. a magnetic rod sleeve; 31. a magnetic attraction pole; 32. a limit ring plate; 33. a puncture section; 34. a rotating gear; 35. an anti-lost ring; 36. a fin wing; 40. a piston rod; 41. a rod body; 42. a first seal ring; 50. a control valve; 51. a valve shaft; 511. a second perforation; 52. an annular soft rubber member; 521. a first perforation; 60. a second seal ring; 70. and a third sealing ring.

Detailed Description

The present invention will be further described with reference to the accompanying drawings and the detailed description, and it should be noted that any combination of the embodiments or technical features described below can be used to form a new embodiment without conflict.

Referring to fig. 1 to 8, a cartridge for rapid nucleic acid detection according to an embodiment of the present invention includes a reagent plate 10, a housing 20, a magnetic rod sleeve 30, a plurality of piston rods 40, and a plurality of control valves 50.

Reagent board 10 and shell 20 swing joint and form the extraction bin that is used for nucleic acid extraction between reagent board 10 and shell 20, reagent board 10 is cylindrical, the reagent board 10 top that is located to extract the bin is concave to be equipped with a plurality of piston channel 11 and a plurality of storage compartment 12, a plurality of piston channel 11 are located the middle part position of reagent board 10, let in the reagent back in storage compartment 12, storage compartment 12 accessible adopts the aluminium membrane heat-seal to seal, the time of adding reaction reagent in testing cycle has been reduced, the periphery in a plurality of piston channel 11 position is located to a plurality of storage compartment 12 rings.

The top of the shell 20 corresponding to the reagent plate 10 is provided with an accommodating cavity 21, a magnetic rod hole 22, a piston hole 23, a sample adding hole 24 and a vent hole 25, the using environment of the rapid nucleic acid detection card box is in an anaerobic environment, oxygen is not required to be completely eliminated, but the oxygen quantity is not too much, therefore, the vent hole 25 can be made very small, certainly, a permeable and impermeable semipermeable membrane can be arranged on the vent hole 25, the vent hole 25 is sealed by adopting the semipermeable membrane in the production process, the semipermeable membrane has the functions of permeable and impermeable liquid, the aperture of the semipermeable membrane is 22nm, the internal and external pressure intensity can be stored in a permeable way, meanwhile, aerosol generated in the magnetic rod extracting sleeve 30 is prevented from leaking, and further, the pollution risk brought by the extracting link is avoided, the sample adding hole 24 only needs to be arranged corresponding to one storage cabin 12 for adding a sample to be detected at least, after the sample is added, the sample adding hole 24 needs to be sealed by a disposable circular airtight pasting sealing film.

It should be noted that the housing 20 can rotate relative to the reagent plate 10 to drive the magnet rod holes 22 to correspond to at least two different storage compartments 12 of the plurality of storage compartments 12, respectively, and the housing 20 can move relative to the reagent plate 10 to change the distance between at least one of the magnet rod holes 22, the sample adding holes 24 and the air holes 25 and the top of the reagent plate 10.

In this embodiment, the piston hole 23 corresponds to the positions of the plurality of piston channels 11, each piston channel 11 is located in the piston hole 23 and is arranged corresponding to the piston hole 23, the magnetic rod hole 22, the sample adding hole 24 and the air vent 25 are all arranged on the periphery of the piston hole 23 and are used for communicating the inside and the outside of the extraction chamber, the magnetic rod hole 22, the sample adding hole 24, the air vent 25 and the piston hole 23 are all communicated with the accommodating cavity 21, and the accommodating cavity 21 is concavely arranged at the bottom of the housing 20; for example, the reagent plate 10 is inserted into the receiving cavity 21, one end of the reagent plate 10 is inserted into the receiving cavity 21, when the reagent plate 10 is moved downward and not separated from the receiving cavity 21, the reagent plate 10 can slide vertically and rotate horizontally relative to the housing 20, and during the sliding and rotating process, the housing 20 and the reagent plate 10 are always kept in a sealed connection to prevent contamination, of course, the housing 20 can also be rotated, and the embodiment is preferably to rotate the reagent plate 10.

The magnetic rod sleeve 30 is rotatably installed on the magnetic rod hole 22 and inserted into one storage cabin 12, one end of the magnetic rod sleeve 30 is located in the extraction cabin, one end of the piston rod 40 is located outside the piston hole 23, the other end of the piston rod passes through the piston hole 23 and then is slidably installed in the piston channel 11 to form a piston structure, the piston channel 11 is communicated with at least one storage cabin 12, in the first embodiment, the piston channel 11 is communicated with one storage cabin 12, the piston channel 11 has a space for providing qPCR amplification and detection, and the control valve 50 is installed on the reagent plate 10 and is used for controlling the connection state between the piston channel 11 and the storage cabin 12 to be connected or disconnected.

The housing 20 is rotatable relative to the reagent plate 10 to rotate at least one of the magnetic rod hole 22, the sample adding hole 24 and the air hole 25 around the piston rod 40, the housing 20 is movable relative to the reagent plate 10 to change the distance between the at least one of the magnetic rod hole 22, the sample adding hole 24 and the air hole 25 and the top of the reagent plate 10, when the reagent plate 10 is descended, the reagent plate 10 and the housing 20 can form a sealed extraction chamber for the magnetic rod sleeve 30 to rotate, the extraction chamber is a sealed structure, the magnetic rod sleeve 30 is in the sealed extraction chamber during the process of ascending along with the housing 20 and entering into the other storage chamber 12, and the magnetic rod sleeve 30 cannot be pulled out from the housing 20.

Referring to fig. 9 to 12, in this embodiment, the reagent plate 10 includes an amplification plate 13 and a base 14 connected to the housing 20, the amplification plate 13 is disposed on the base 14, the base 14 is inserted into the accommodating cavity 21 of the housing 20, the number of the piston channels 11 is one, the middle portion of the base 14 is sequentially communicated with a guide hole 141 and a mounting hole 142 from top to bottom, the storage chamber 12 is recessed on the base 14, the amplification plate 13 is mounted on the mounting hole 142 from the bottom of the base 14, an amplification channel 111 is disposed in the amplification plate 13, the amplification channel 111 has the space for amplification and detection, the base 14 is further provided with a micro channel 143 communicated with the storage chamber 12, one end of the amplification channel 111 is communicated with the guide hole 141, the other end is communicated with the micro channel 143, and the guide hole 141, the amplification channel 111 and the microchannel 143 constitute the piston channel 11, and the control valve 50 is mounted on the base 14 and controls the opening and closing of the microchannel 143.

The amplification pipeline 111 comprises a guide channel 1111, a first liquid guide pipe 1112, an amplification chamber 1113 and a second liquid guide pipe 1114 which are sequentially communicated, the volume of the amplification chamber 1113 is larger than the volume of the first liquid guide pipe 1112 and the volume of the second liquid guide pipe 1114, the amplification chamber 1113 is used for providing amplification and detection space for qPCR amplification detection, the guide channel 1111 is communicated with the guide hole 141, and the second liquid guide pipe 1114 is communicated with the micro-channel 143.

The plurality of storage compartments 12 include a first chamber 121 for storing the lysis binding solution, a second chamber 122 for storing the first washing solution, a third chamber 123 for storing the second washing solution, a fourth chamber 124 for storing the eluent, a fifth chamber 125 for storing the magnetic rod sleeve 30, and a sixth chamber 126 for storing the magnetic beads, the first chamber 121, the second chamber 122, the third chamber 123, the fourth chamber 124, the fifth chamber 125, and the sixth chamber 126 are separately disposed, and the fourth chamber 124 communicates with the microchannel 143, so that the reagent plate 10 is also referred to as a six-well reagent plate, and the first chamber 121 is also referred to as a sample processing chamber.

The first chamber 121 can contain 5mL of reagent at most, and the first chamber 121 mainly contains 2.5mL of lysis binding solution, which mainly has the effects of destroying the sample structure and releasing nucleic acid;

the second chamber 122 can contain 2.2mL of reagent at most, and the second chamber 122 mainly contains 0.6mL of the first washing solution, and the main function is to wash the heteroprotein residues on the magnetic beads;

the third chamber 123 can contain 2.2mL of reagent at most, and the third chamber 123 mainly contains 0.6mL of second washing solution which has the main function of washing the salt ion residue on the magnetic beads;

the fourth chamber 124 can contain 1.0mL of reagent at most, the fourth chamber 124 mainly contains 100 μ L of eluent, and the main function is to elute the nucleic acid molecules captured on the magnetic beads into the eluent to obtain an extraction product;

the sixth chamber 126 can contain 2.2mL of reagent at most, and the sixth chamber 126 mainly contains 200 μ L of magnetic bead solution, and the magnetic beads mainly function to specifically adsorb nucleic acid under the environment of high salt and low pH, i.e. to adsorb nucleic acid on the magnetic beads, and release nucleic acid under the environment of low salt and high pH, i.e. to separate magnetic beads from nucleic acid.

The amplification plate 13 comprises a columnar main body 131, a fixed ring plate 132 and a flat body 133 positioned at the bottom of the columnar main body 131, the fixed ring plate 132 is convexly arranged on the outer side wall of the columnar main body 131, the reagent plate 10 further comprises a fourth sealing ring 15, the columnar main body 131 is positioned in the mounting hole 142, the fourth sealing ring 15 is clamped between the inner wall of the mounting hole 142 and the outer wall of the columnar main body 131 to form sealing, the fixed ring plate 132 abuts against the bottom of the reagent plate 10, the pre-packaged freeze-dried amplification reagent is stored in the amplification chamber 1113, the other more preferable design is that the pre-packaged freeze-dried amplification reagent is stored in the guide channel 1111, and the fixed ring plate 132 plays a role in stably fixing the amplification plate 13; the amplification chamber 1113 in the flat body 133 is designed smoothly, has an internal volume of 50-60 μ L, and is mainly used for storing a freeze-drying amplification reagent and subsequently performing a real-time fluorescence polymerase chain reaction.

Referring to fig. 13 and 15, the bottom of the base 14 is provided with one valve through hole 144, the number of the control valves 50 is one, the micro fluid channel 143 includes a first micro fluid pipe 1431 communicated with the second fluid pipe 1114 and a second micro fluid pipe 1432 communicated with the fourth chamber 124, the valve through hole 144 divides the micro fluid channel 143 into two sections, the control valve 50 is a one-way valve, the control valve 50 is installed on the valve through hole 144, and one end of the control valve 50 is communicated with the first micro fluid pipe 1431 and the other end is communicated with the second micro fluid pipe 1432.

The control valve 50 comprises a valve shaft 51 and a ring-shaped soft rubber 52, the ring-shaped soft rubber 52 is sleeved on the valve shaft 51, the ring-shaped soft rubber 52 and the valve shaft 51 are respectively provided with a first perforation 521 and a second perforation 511, the valve shaft 51 and the ring-shaped soft rubber 52 are inserted into the valve through hole 144, when the first perforation 521, the second perforation 511 and the micro-channel 143 are communicated, the control valve 50 is in an open state, otherwise, the control valve 50 is in a closed state during the nucleic acid extraction operation, the connection between the fourth chamber 124 and the pre-packaged freeze-dried amplification reagent in the amplification pipeline 111 is cut off, when the nucleic acid extraction is completed, the control valve 50 realizes pipeline communication through rotation, the extracted nucleic acid molecules flow into the amplification chamber 1113 through the pressure generated by the piston rod 40 in the piston channel 11 to dissolve and pre-mix the freeze-dried amplification reagent, the opening and closing of the pipeline are realized through rotating the control valve 50, the piston rod 40 is matched with the amplification pipeline 111 to realize the transfer of the eluent into the amplification pipeline 111.

The piston rod 40 includes a rod body 41 and a first sealing ring 42, the outer diameter of the rod body 41 is smaller than the inner diameter of the piston channel 11, the rod body 41 is inserted into the piston channel 11, an annular first mounting groove is concavely arranged on the outer side wall of the rod body 41, the inner end of the first sealing ring 42 is mounted in the first mounting groove, the outer end of the first sealing ring 42 abuts against the inner wall of the piston channel 11, the front end of the piston rod 40 is inserted into the piston channel 11 through the first sealing ring 42 on the front end of the piston rod to realize sealing, a detection instrument grasps the top end of the piston rod 40, and the inner and outer pressure can be changed through the movement of the piston rod 40 to serve as a power source of the driving liquid.

The top of the reagent plate 10 is further concavely provided with an annular cavity 16, the piston channel 11 and the storage chamber 12 are respectively located at the inner side and the outer side of the annular cavity 16, the housing 20 comprises an annular top plate 26, an annular enclosing plate 27 bent and extending from the outer edge of the annular top plate 26, and a positioning tube 28 located in the annular enclosing plate 27 and coaxially arranged with the annular enclosing plate 27, the annular enclosing plate 27 and the positioning tube 28 are both of tube body structures, a piston hole 23, a magnetic rod hole 22, the sampling hole 24 and the air hole 25 are both arranged on the annular top plate 26, the positioning tube 28 and the annular enclosing plate 27 are both arranged at the bottom of the annular top plate 26 in a protruding mode, the positioning tube 28 is connected with the annular top plate 26, the inner diameter of the positioning tube 28 is larger than that of the piston hole 23, the magnetic rod hole 22, the sampling hole 24 and the air hole 25 are all located between the positioning tube 28 and the annular enclosing plate 27, the positioning tube 28 is inserted into the annular cavity 16, and the bottom wall of the annular top plate 26 and the inner wall of the annular enclosing plate 27 are enclosed to form the accommodating cavity 21.

As a preferred embodiment, the cartridge for rapid nucleic acid detection further comprises a second sealing ring 60, wherein an annular second mounting groove is concavely formed on the outer side wall of the reagent plate 10, the second mounting groove is arranged near the annular top plate 26, the inner end of the second sealing ring 60 is mounted in the second mounting groove, and the outer end is in sliding fit with the annular enclosing plate 27.

As a preferred embodiment, the cartridge for rapid detection of nucleic acid further comprises a third sealing ring 70, an annular third mounting groove is concavely provided on the outer side wall of the positioning tube 28, the third mounting groove is disposed away from the annular top plate 26, the inner end of the third sealing ring 70 is mounted in the third mounting groove, the outer end is in sliding fit with the inner wall of the annular cavity 16 to form a closed nucleic acid extraction chamber as described above, the piston hole 23 is not connected with the extraction chamber and is mainly used for the piston rod 40 to pass through, and the first sealing ring 42, the second sealing ring 60 and the third sealing ring 70 mainly play a role of sealing the extraction chamber.

The magnetic rod sleeve 30 comprises a magnetic suction rod 31, a limit ring plate 32 and a puncture part 33, the puncture part 33 is convexly arranged at the bottom of the magnetic suction rod 31, the limit ring plate 32 is convexly arranged on the outer side wall of the magnetic suction rod 31, the limit ring plate 32 is rotatably arranged on the magnetic rod hole 22, and one end of the magnetic suction rod 31, which is provided with the puncture part 33, is inserted into the storage chamber 12, in the embodiment, the magnetic rod sleeve 30 needs to puncture each aluminum film on the reagent plate 10 to enter each storage chamber 12 respectively, so that the conical puncture part 33 is designed at the bottom end of the magnetic rod sleeve 30 to puncture the aluminum films, and the limit ring plate 32 is matched with the magnetic rod hole 22 to ensure that the magnetic rod sleeve 30 is fixed on the circular shell 20 and the magnetic rod sleeve 30 can only rotate on a horizontal plane; meanwhile, the top of the magnetic attraction rod 31 is provided with a rotating gear 34, and the rotating gear 34 can be matched with a detection instrument to realize the rotation of the magnetic rod sleeve 30, so that the cleaning of magnetic beads and the uniform mixing of reagents are realized.

As a preferred embodiment, in order to prevent the magnetic beads on the magnetic rod sleeve 30 from being scratched by the aluminum film and lost during the process of puncturing the aluminum film and removing the magnetic rod sleeve 30 from the storage compartment 12, the magnetic rod sleeve 30 of the present invention further includes an anti-lost ring 35, the anti-lost ring 35 is in a ring shape, the anti-lost ring 35 is convexly disposed at the bottom of the magnetic attraction rod 31, the inner diameter of the anti-lost ring 35 gradually decreases from one end away from the magnetic attraction rod 31 to one end close to the magnetic attraction rod 31, the puncture part 33 is disposed in the anti-lost ring 35, the axial distance from one end of the anti-lost ring 35 away from the magnetic attraction rod 31 to the magnetic attraction rod 31 is smaller than the axial distance from one end of the puncture part 33 away from the magnetic attraction rod 31 to the magnetic attraction rod 31, when the magnetic rod sleeve 30 adsorbs the magnetic beads, the magnetic beads are adsorbed into the groove between the anti-lost ring 35 and the puncture part 33, or in a groove above the anti-lost ring 35, so that the magnetic beads cannot be in direct contact with the aluminum film, thereby reducing the loss of the magnetic beads.

As a preferred embodiment, the magnetic rod sleeve 30 further includes a plurality of fin wings 36, the fin wings 36 are convexly disposed on the outer side wall of the magnetic suction rod 31, the fin wings 36 are located between the limit ring plate 32 and the anti-lost ring 35 and are disposed close to the anti-lost ring 35, the magnetic rod sleeve 30 of the present invention adopts a rotary design, and the stirring resistance is increased when the fin wings 36 are added for stirring the reagent, so as to achieve rapid mixing of the reagent.

Referring to fig. 16 and 17, a second embodiment of the present invention also discloses a rapid nucleic acid detecting cartridge, which is different from the first embodiment in that the structure of the reagent plate 10 is different, the reagent plate 10 in the second embodiment includes a base 14 and an amplification plate 13, the base 14 is provided with a plurality of storage compartments 12, the plurality of storage compartments 12 includes a first chamber 121 for storing a lysis binding solution, a second chamber 122 for storing a first washing solution, a third chamber 123 for storing a second washing solution, a fourth chamber 124 for storing an eluent, a fifth chamber 125 for storing a magnetic rod sleeve 30, a sixth chamber 126 for storing magnetic beads, and a seventh chamber 127 for storing an amplification freeze-drying reagent, the first chamber 121, the second chamber 122, the third chamber 123, the fifth chamber 125, and the sixth chamber 126 are separately disposed, the fourth chamber 124 and the seventh chamber 127 are both communicated with a micro flow channel 143, the freeze-dried amplification reagents include hot-start Taq enzyme, dNTPs, primers, probes and the like, and therefore, the reagent plate 10 in the second embodiment is also referred to as a seven-well reagent plate.

In the second embodiment, compared with the first embodiment, a seventh chamber 127 is added for storing the amplification freeze-drying reagent separately, and the micro flow channel 143 is redesigned, in the present embodiment, the number of the control valves 50 is one, the micro flow channel 143 is designed in a "Y" shape with an angle of 120 °, the micro flow channel 143 includes a third micro flow tube communicated with the second liquid guide tube 1114, a fourth micro flow tube communicated with the fourth chamber 124, and a fifth micro flow tube communicated with the seventh chamber 127, the control valves 50 are three-way valves, one end of the control valves 50 is communicated with the third micro flow tube, and the other end is communicated with the fourth micro flow tube and the fifth micro flow channel 143, respectively, and the control valves 50 are disposed at the intersection of the Y-shaped micro flow channel 143.

Of course, in another embodiment, the number of the control valves 50 is two, the micro flow channel 143 is "Y" shaped, the micro flow channel 143 includes a third micro flow tube connected to the second fluid conduit 1114, a fourth micro flow tube connected to the fourth chamber 124, and a fifth micro flow tube connected to the seventh chamber 127, the fourth micro flow tube and the fifth micro flow tube are both connected to the third micro flow tube, both of the control valves 50 are one-way valves, one of the control valves 50 is used to control the on/off of the fourth micro flow tube, and the other control valve 50 is used to control the on/off of the fifth micro flow tube.

The optimized design is suitable for developing nested qPCR amplification, the amplification product is pushed into the seventh chamber 127 after the first round of qPCR amplification is completed in the pre-amplification pipeline 111, and after reagents are uniformly mixed, the second round of qPCR amplification can be developed, so that the detection sensitivity can be further improved.

The invention also provides a method for rapidly detecting nucleic acid, which comprises the above-mentioned cassette for rapidly detecting nucleic acid, wherein each reagent is respectively stored in each storage cabin 12 and amplification cabin 1113, each storage cabin 12 and amplification cabin 1113 are in a sealed state, and the control valve 50 is in a closed state, taking six-hole reagent plates as an example, the cassette for rapidly detecting nucleic acid can be also called as a cassette for short as follows, and comprises the following steps:

step a, adding 50 mu L of proteinase K solution and a sample to be detected into a first cavity 121 through a sample adding hole 24 in sequence by using a liquid transfer machine, blowing and uniformly mixing for 5-8 times by using the liquid transfer machine, sealing the sample adding hole 24, then placing a card box into a specified position of a detection instrument according to the indication of the card box, starting a software operation interface of the detection instrument, selecting a corresponding detection item program, and starting a detection flow, wherein the detection instrument has the functions of lifting a reagent plate 10, rotating the reagent plate 10, a magnetic rod sleeve 30 and the like.

B, using a detection instrument to lower the circular reagent plate 10 to a certain height, lifting the reagent plate 10 to form a sealed extraction bin, rotating the circular reagent plate 10 clockwise through the extraction bin, positioning the magnetic rod sleeve 30 above the first chamber 121, then lifting the circular reagent plate 10 to a specified height, inserting the magnetic rod sleeve 30 into the first chamber 121 to stir, using the detection instrument to rotate the magnetic rod sleeve 30, uniformly mixing the sample cracking binding solution at a certain rotating speed, and using a heating module on the detection instrument to heat the liquid in the first chamber 121, so that the sample to be detected is cracked to release nucleic acid;

step c, adsorbing the magnetic beads in the sixth chamber 126 into the first chamber 121 by using the magnetic rod sleeve 30 through the extraction chamber, specifically, lowering the circular reagent plate 10 to a certain height by using a detection instrument, rotating the circular reagent plate 10 counterclockwise by 60 degrees to enable the magnetic rod sleeve 30 to be positioned above the sixth chamber 126, then raising the circular reagent plate 10 to a specified height, inserting the magnetic rod sleeve 30 into the sixth chamber 126, lowering the magnetic rod sleeve 30 to a certain position to adsorb the magnetic beads in the sixth chamber 126 for 1min, and adsorbing the magnetic beads on the magnetic rod sleeve 30;

the method comprises the following steps that a detection instrument is utilized to lower a circular reagent plate 10 to a certain height, the circular reagent plate 10 is rotated clockwise by 60 degrees through an extraction bin, a magnetic rod sleeve 30 is located above a first chamber 121, then the circular reagent plate 10 is lifted to a specified height, the magnetic rod sleeve 30 is inserted into the first chamber 121 at the moment, magnetic beads in a sixth chamber 126 are adsorbed into the first chamber 121, the magnetic rod sleeve 30 is driven to enable the magnetic rod sleeve 30 to uniformly mix a sample splitting and combining solution at a certain rotating speed, the mixing time is 4min, the magnetic beads adsorb nucleic acid at the moment, but can adsorb impurities such as protein at the same time, the magnetic rod sleeve 30 is lowered to a certain position, the magnetic beads with the nucleic acid are adsorbed on the magnetic rod sleeve 30, and the impurities such as the protein are contained in the magnetic beads at the moment;

step d, adsorbing the magnetic beads with the nucleic acid in the first chamber 121 to the second chamber 122 through the extraction chamber, removing impure proteins on the magnetic beads, specifically, lowering the circular reagent plate 10 to a certain height, sucking the magnetic beads with nucleic acids out of the first chamber 121, rotating the circular reagent plate 10 clockwise by 60 degrees, so that the magnetic rod sleeve 30 is positioned above the second chamber 122, the circular reagent plate 10 is then raised to a specified height, at which time the magnetic sleeve 30 is inserted into the second chamber 122, after the magnetic rod sleeve 30 is quickly lifted to a certain position, the instrument is started to uniformly mix the sample lysis binding solution by the magnetic rod sleeve 30 through the rotating gear 34 on the magnetic rod sleeve 30 at a certain rotating speed for 1min, the magnetic beads with nucleic acid are adsorbed into the second chamber 122, removing the impure proteins on the magnetic beads, wherein the first washing solution in the second chamber 122 is an ethanol solution of guanidine salt;

step e, adsorbing the magnetic beads with the nucleic acids in the second chamber 122 into the third chamber 123 through the extraction chamber, and removing salt ions on the magnetic beads, specifically, lowering the circular reagent plate 10 to a certain height by using a detection instrument, rotating the circular reagent plate 10 clockwise by 60 degrees through the extraction chamber, so that the magnetic rod sleeve 30 is positioned above the third chamber 123, then raising the circular reagent plate 10 to a specified height, inserting the magnetic rod sleeve 30 into the third chamber 123 at this time, adsorbing the magnetic beads with the nucleic acids in the second chamber 122 into the third chamber 123, rapidly raising the magnetic rod sleeve 30 to a certain position, starting the instrument to rotate the magnetic rod sleeve 30, so that the magnetic rod sleeve 30 uniformly mixes the sample solution at a certain rotation speed for 1min, and removing the salt ions on the magnetic beads, wherein the second washing solution in the third chamber 123 is 70% or 80% ethanol solution;

step f, lowering the circular reagent plate 10 to a certain height, rotating the circular reagent plate 10 clockwise by 60 degrees through the extraction chamber to enable the magnetic rod sleeve 30 to be positioned above the fourth chamber 124, standing for 2min, then raising the circular reagent plate 10 to a specified height, inserting the magnetic rod sleeve 30 into the fourth chamber 124, and adsorbing magnetic beads with nucleic acid in the third chamber 123 into the fourth chamber 124;

after the magnetic rod sleeve 30 is quickly lifted to a certain position, the instrument starts the magnetic rod sleeve 30 to rotate, so that the magnetic rod sleeve 30 uniformly mixes the sample solution according to a certain rotating speed, meanwhile, the heating module is lifted to a position below the fourth chamber 124, the temperature is raised to 75 +/-1 ℃, the mixing time is 3min, and the nucleic acid molecules captured on the magnetic beads are eluted by using the elution liquid in the fourth chamber 124;

through the extraction bin, the circular reagent plate 10 is lowered to a certain height, then the magnetic rod is lowered to a certain position for 1min to only adsorb magnetic beads, then the circular reagent plate 10 is rotated clockwise by 60 degrees, the magnetic rod sleeve 30 is positioned above the fifth chamber 125, then the circular reagent plate 10 is raised to a specified height, at this time, the magnetic rod sleeve 30 is inserted into the fifth chamber 125, the magnetic rod is quickly raised to a certain position, the magnetic beads are removed from the fourth chamber 124, and the magnetic beads are specifically transferred into the fifth chamber 125;

step g, opening the control valve 50, rotating the valve shaft 51 to enable the micro flow channel 143 to be in a communication state, slowly pulling up the piston rod 40 to adsorb the nucleic acid molecules in the fourth chamber 124 into the amplification chamber 1113, dissolving the pre-packaged amplification freeze-drying reagent in the amplification chamber 1113, and uniformly mixing;

and closing the control valve 50 to enable the micro flow channel 143 to be in a closed state, starting the temperature control module and the optical detection module to control an amplification program of the amplification tube, starting the detection instrument to automatically perform qPCR amplification detection, enabling the nucleic acid to react with the amplification freeze-drying reagent, automatically collecting a fluorescence signal in each cycle reaction in the amplification chamber 1113, displaying the collected signal on a software interface in real time after optimization processing, completing fluorescence curve analysis after 30min, and automatically giving an analysis result by the instrument and prompting an operator to give a detection report.

And clicking on a software interface to complete an experiment, automatically ejecting the nucleic acid rapid detection card box by the instrument, taking the nucleic acid rapid detection card box out, putting the nucleic acid rapid detection card box into a self-sealing bag to perform corresponding treatment according to medical instrument garbage, and then turning off a power supply or turning on the next round of detection.

The specific steps of storing the reagents in the storage chambers 12 and the amplification chamber 1113 respectively, wherein the storage chambers 12 and the amplification chamber 1113 are in a sealed state are as follows:

detaching the reagent plate 10, storing the lysis binding solution in the first chamber 121 and heat-sealing with an aluminum film, storing the first wash solution in the second chamber 122 and heat-sealing with an aluminum film, storing the second wash solution in the third chamber 123 and heat-sealing with an aluminum film, storing the eluate in the fourth chamber 124 and heat-sealing with an aluminum film, storing magnetic beads in the sixth chamber 126 and heat-sealing with an aluminum film, storing the amplification freeze-drying reagent in the amplification chamber 1113 and sealing with the piston rod 40 and the control valve 50;

the reagent plate 10 is inserted into the receiving cavity 21 of the housing 20.

In step b, the temperature of the liquid in the first chamber 121 is raised to 75 ± 1 ℃ and kept for 5 minutes.

The specific embodiment is as follows: the detection reagent adopts influenza A virus specificity detection reagent, the actions of nucleic acid extraction of 1mL of pseudovirus simulation sample, the even mixing of amplification reagent and the like are manually completed according to the operation steps, then 50 muL of system is absorbed from the evenly mixed amplification reagent and is put into ABI7500 for fluorescence quantitative qPCR amplification, meanwhile, the nucleic acid of 200 muL of pseudovirus simulation sample is extracted by adopting the conventional virus nucleic acid extraction reagent, 100 muL of eluent is adopted for elution, 10 muL of extraction product is absorbed and is taken as a template, 50 muL of amplification system is put into ABI7500 for amplification, the amplification result is taken as a contrast, the amplification result is shown in figure 18, the result shows that the nucleic acid rapid detection card box designed by the invention can be detected 4-5 cycles in advance, the detection rate is obviously improved for low-abundance samples, and the detection sensitivity is far higher than that of the conventional detection method, in FIG. 18, the single-dot chain line is for the present invention and the two-dot chain line is for the conventional qPCR amplification data.

The invention has the following advantages:

1. in order to realize the purpose of closed nucleic acid extraction and purification, the invention innovatively adopts the design of the disc type reagent plate 10 and the design of the closed shell 20, and realizes the nucleic acid extraction and purification of the magnetic rod sleeve 30 in the closed card box for the first time through the innovation, thereby solving the pollution problem caused by aerosol formed in the nucleic acid extraction process;

2. in order to realize the purpose of closed nucleic acid extraction and purification, the invention adopts the design of the fin-wing 36-type magnetic rod sleeve 30, and the uniform mixing of reagents is realized through rotation, so that aerosol generated by severe vibration can be avoided, and the closure of the card box is kept;

3. in order to avoid the loss of the magnetic beads on the magnetic rod sleeve 30 in the extraction process, the invention creatively designs the anti-lost ring 35 of the magnetic rod sleeve 30 to ensure that the magnetic beads are not lost due to the contact of the sealing film when the magnetic beads puncture the sealing film and leave the storage cabin 12;

4. in order to realize the performance index of ultra-high detection limit, the volume of the first chamber 121 is innovatively designed, the nucleic acid extraction with the sample volume of 1mL can be realized, and compared with the design of other molecule POCTs on the market, the invention has the advantage of extracting large-volume samples by operation, thereby greatly improving the detection rate;

5. in order to realize the performance index of ultra-high detection limit, the amplification pipeline 111 is innovatively integrated on the circular reagent plate 10, and the length of the micro-channel 143 is greatly reduced, so that the template volume ratio in an amplification system is greatly improved, and the minimum detection limit is further improved.

6. The nucleic acid rapid detection card box has ultrahigh detection sensitivity, 1mL of clinical sample is put in, 100 mu L of eluent is used for eluting and extracting products, wherein not less than 50 mu L of clinical sample is put in an amplification system, so that the lowest detection limit of 10 copies/mL can be realized, and the invention has absolute leading advantage on the lowest detection limit for the traditional detection reagent in the market or other molecular POCT products.

7. The rapid nucleic acid detection card box has fewer components and low processing difficulty, can realize automation in production and assembly, improves the production efficiency, greatly reduces the production cost, and has a leading advantage in cost compared with other solutions in the market.

8. The nucleic acid rapid detection card box provided by the invention can be applied to various hospitals, disease control centers, blood stations, mobile blood donation stations, customs and pet hospitals, and can also be applied to other various field detection scenes, including rural clinics in remote areas, outdoor environment monitoring and the like.

The above embodiments are only preferred embodiments of the present invention, and the protection scope of the present invention is not limited thereby, and any insubstantial changes and substitutions made by those skilled in the art based on the present invention are within the protection scope of the present invention.

Claims

1. The utility model provides a quick detection card box of nucleic acid, its characterized in that, includes reagent board, shell, bar magnet cover, a plurality of piston rod and a plurality of control valve, the reagent board with shell swing joint and form the extraction bin that is used for nucleic acid to draw between reagent board and the shell, be located draw the bin in the concave a plurality of piston passageways and a plurality of storage compartment that are equipped with in top of reagent board, a plurality of the storage compartment ring is located the periphery of piston passageway position, correspond on the shell the top of reagent board has bar magnet hole, application of sample hole, bleeder vent and corresponds the piston hole of a plurality of piston passageway position, bar magnet hole, application of sample hole and bleeder vent are located the periphery of piston hole and are used for the intercommunication draw bin inside and outside, the bar magnet cover rotate install in on the bar magnet hole and be located draw the bin is indoor, the one end of piston rod is located outside the piston hole, the other end pass behind the piston hole slidable mounting in the piston passageway, the shell can for the reagent board rotates in order to drive the bar magnet hole respectively correspond to two at least different storage cabins in a plurality of storage cabins, the shell can for the reagent board removes in order to change at least one in bar magnet hole, the hole of adding the appearance and the bleeder vent with the distance between the top of reagent board, piston passageway and at least one the storage cabin intercommunication, piston passageway has the space that is used for augmenting and detecting, the control valve install in on the reagent board and be used for controlling the piston passageway with the break-make of intercommunication state between the storage cabin.

2. The rapid nucleic acid detecting cartridge according to claim 1, wherein: the reagent board is convexly provided with an amplification board, an amplification pipeline is arranged in the amplification board, the piston channel comprises the amplification pipeline, and the amplification pipeline is provided with a space for amplification and detection.

3. The rapid nucleic acid detecting cartridge according to claim 2, wherein: the reagent board still have with the base that the shell is connected, the amplification board sets up on the base, the quantity of piston passageway is one, the middle part top-down of base is equipped with guiding hole and mounting hole in proper order, the storage cabin is concave to be located on the base, the amplification board install in on the mounting hole, still be equipped with on the base with the microchannel of storage cabin intercommunication, the one end of amplification pipeline with guiding hole intercommunication, the other end with the microchannel intercommunication, guiding hole, amplification pipeline and microchannel constitute piston passageway, the control valve install in on the base and be used for controlling the break-make of microchannel.

4. The rapid nucleic acid detecting cartridge according to claim 3, wherein: the amplification pipeline comprises a guide channel, a first liquid guide pipe, an amplification chamber and a second liquid guide pipe which are sequentially communicated, the amplification chamber is used for providing the space for amplification and detection, the guide channel is communicated with the guide hole, and the second liquid guide pipe is communicated with the micro-channel.

5. The rapid nucleic acid detecting cartridge according to claim 4, wherein: the storage cabin comprises a first chamber for storing the split binding liquid, a second chamber for storing the first washing liquid, a third chamber for storing the second washing liquid, a fourth chamber for storing the eluent, a fifth chamber for storing the magnetic rod sleeve and a sixth chamber for storing the magnetic beads, wherein the first chamber, the second chamber, the third chamber, the fourth chamber, the fifth chamber and the sixth chamber are arranged in an isolated mode, and the fourth chamber is communicated with the micro-channel.

6. The rapid nucleic acid detecting cartridge according to claim 5, wherein: the number of the control valves is one, the micro flow channel comprises a first micro flow pipe communicated with the second liquid guide pipe and a second micro flow pipe communicated with the fourth cavity, the control valves are one-way valves, one end of each control valve is communicated with the first micro flow pipe, and the other end of each control valve is communicated with the second micro flow pipe.

7. The rapid nucleic acid detecting cartridge according to claim 4, wherein: the storage cabin comprises a first chamber for storing the split binding liquid, a second chamber for storing the first washing liquid, a third chamber for storing the second washing liquid, a fourth chamber for storing the eluent, a fifth chamber for storing the magnetic rod sleeve, a sixth chamber for storing the magnetic beads and a seventh chamber for storing the amplification freeze-drying reagent, wherein the first chamber, the second chamber, the third chamber, the fifth chamber and the sixth chamber are arranged in an isolated mode, and the fourth chamber and the seventh chamber are communicated with the micro-channel.

8. The rapid nucleic acid detecting cartridge according to claim 7, wherein: the number of the control valve is one, the micro flow channel comprises a third micro flow pipe communicated with the second liquid guide pipe, a fourth micro flow pipe communicated with the fourth cavity and a fifth micro flow pipe communicated with the seventh cavity, the control valve is a three-way valve, one end of the control valve is communicated with the third micro flow pipe, and the other end of the control valve is communicated with the fourth micro flow pipe and the fifth micro flow channel respectively.

9. The rapid nucleic acid detecting cartridge according to claim 7, wherein: the quantity of control valve is two, the miniflow channel include with the third miniflow pipe of second catheter intercommunication, with the fourth miniflow pipe of fourth cavity intercommunication and with the fifth miniflow pipe of seventh cavity intercommunication, fourth miniflow pipe and fifth miniflow pipe all with third miniflow pipe intercommunication, two the control valve all is the check valve, one the control valve is used for controlling the break-make of fourth miniflow pipe, another the control valve is used for controlling the break-make of fifth miniflow pipe.

10. The rapid nucleic acid detecting cartridge according to claim 1, wherein: the piston rod comprises a rod body and a first sealing ring, the rod body is inserted into the piston channel, an annular first mounting groove is concavely arranged on the outer side wall of the rod body, and the inner end of the first sealing ring is mounted in the first mounting groove and the outer end of the first sealing ring is abutted to the inner wall of the piston channel.

11. The rapid nucleic acid detecting cartridge according to claim 1, wherein: the bottom of shell is equipped with in a recess and holds the chamber, reagent board card insert in hold the intracavity, bar magnet hole, application of sample hole, bleeder vent and piston hole all with hold the chamber intercommunication.

12. The rapid nucleic acid detecting cartridge according to claim 11, wherein: the top of reagent board is the concave annular chamber that is equipped with still, piston passageway and storage chamber are located respectively the inside and outside both sides of annular chamber, the shell includes annular roof, certainly the outward flange of annular roof is buckled the annular bounding wall that extends and is located in the annular bounding wall and with the registration arm of the coaxial setting of annular bounding wall, piston hole, bar hole, application of sample hole and bleeder vent are all located on the annular roof, the registration arm with the annular roof is connected, the internal diameter of registration arm is greater than the internal diameter of piston hole, bar hole, application of sample hole and bleeder vent all are located between registration arm and the annular bounding wall, the registration arm insert in the annular chamber, the diapire of annular roof and the inner wall of annular bounding wall enclose to be synthetic hold the chamber.

13. The rapid nucleic acid detecting cartridge according to claim 12, wherein: the quick nucleic acid detection card box further comprises a second sealing ring, an annular second mounting groove is concavely formed in the outer side wall of the reagent plate, the second mounting groove is close to the annular top plate, and the inner end of the second sealing ring is mounted in the second mounting groove, the outer end of the second sealing ring is in sliding fit with the annular enclosing plate.

14. The rapid nucleic acid detecting cartridge according to claim 12, wherein: the quick nucleic acid detection card box further comprises a third sealing ring, an annular third mounting groove is concavely formed in the outer side wall of the positioning tube, the third mounting groove is far away from the annular top plate, and the inner end of the third sealing ring is mounted in the third mounting groove, the outer end of the third sealing ring is in sliding fit with the inner wall of the annular cavity.

15. The rapid nucleic acid detecting cartridge according to claim 1, wherein: the bar magnet cover includes that the magnetism inhales pole, spacing ring board and puncture portion, the protruding bottom of inhaling the pole of puncture portion of locating of magnetism, the protruding locating of spacing ring board on the lateral wall of inhaling the pole of magnetism, spacing ring board rotate install in on the bar magnet hole, the one end that has of magnetism inhale the pole insert in the storage compartment, the bar magnet cover is still including preventing losing the circle, it is cyclic annular to prevent losing the circle, prevent losing the protruding locating of circle the bottom of magnetism inhaling the pole, prevent losing the internal diameter of circle from keeping away from the one end of magnetism inhaling the pole is close to one end of magnetism inhaling the pole reduces gradually, the puncture position in prevent losing the circle, prevent losing keeping away from of circle the one end of magnetism inhaling the pole extremely the axial distance of magnetism inhaling the pole is less than the axial distance of puncture portion keep away from the one end of magnetism inhaling the pole extremely the axial distance of magnetism inhales the pole.

16. The rapid nucleic acid detecting cartridge according to claim 15, wherein: the magnetic rod sleeve further comprises a plurality of fin wings, the fin wings are convexly arranged on the outer side wall of the magnetic suction rod, and the fin wings are located between the limiting ring plate and the anti-lost ring and are close to the anti-lost ring.

17. The rapid nucleic acid detecting cartridge according to any one of claims 1 to 16, wherein: and a permeable and impermeable semipermeable membrane is arranged on the air holes.

18. A method for rapid detection of nucleic acid comprising the nucleic acid rapid detection cartridge according to claim 5, wherein each reagent is stored in each storage compartment and each amplification compartment, and each storage compartment and each amplification compartment are sealed, and the control valve is in a closed state, the method comprising:

19. The method for rapid detection of nucleic acid according to claim 18, wherein: the method comprises the following steps of respectively storing each reagent in each storage cabin and each amplification cabin, wherein the storage cabins and the amplification cabins are in a sealed state:

20. The method for rapid detection of nucleic acid according to claim 18, wherein: and a permeable and impermeable semipermeable membrane is arranged on the air holes.