A nucleic acid detection card box and its assembly process

Abstract

A nucleic acid detection card box includes a sampling tube and a box body. The box body includes a box body and an upper cover plate and a lower cover plate respectively covering the upper and lower parts of the box body. A plurality of cavities for containing substances used for nucleic acid extraction are arranged in the box body, and the sampling tube is connected to one of the cavities; the upper cover plate is provided with corresponding feed holes, and the lower cover plate is bonded with the box body to form fluid channels respectively connected to the cavities, and the fluid channels can be selectively connected. The fully enclosed card box structure of the present invention is well designed, low-cost, extremely simple to operate, and can realize the integration of extraction, amplification and detection. It is suitable for various scenarios, from the emergency department of a tertiary hospital to the community and township health center, and is particularly suitable for environments where conventional instruments are not suitable for work.

Description

Technical Field

[0001] The invention relates to a nucleic acid extraction technology, and in particular to a nucleic acid detection card box and a corresponding assembly process. Background Art

[0002] As a hot direction for the development of molecular diagnosis in the future, molecular diagnosis POCT combines the characteristics of being small, flexible, convenient, fast, and accurate. It has more and wider application scenarios and fields than traditional molecular diagnosis. This is also the reason why domestic and foreign diagnostic companies are deploying in this field. However, to achieve the true meaning of "sample in, result out" molecular diagnosis POCT requires the following basic elements: fully enclosed cartridge design, integrated extraction + amplification + detection, extremely simple operation, low cost, multiplex PCR technology, reagent room temperature technology, rapid extraction and purification technology, and portable molecular diagnosis POCT instrument. At present, there are no molecular POCT products on the market that truly realize the above basic elements. Summary of the invention

[0003] In order to solve the deficiencies in the prior art, the present invention provides a nucleic acid detection card box and its assembly process, with a sophisticatedly designed fully enclosed card box structure, low cost, and extremely simple operation, which can realize the integration of extraction, amplification and detection.

[0004] To achieve the above object, the present invention provides the following technical solutions:

[0005] A nucleic acid detection card box comprises a sampling tube and a box body. The box body is provided with a plurality of cavities for containing substances used for nucleic acid extraction and a plurality of fluid channels connected to the cavities. The sampling tube is connected to one of the cavities, and the fluid channels can be selectively connected.

[0006] Furthermore, the box body includes a box body and an upper cover plate covering the box body, the upper cover plate is provided with a feed hole corresponding to the cavity, and the upper end surface of the upper cover plate is covered with a top sealing film for sealing.

[0007] Preferably, the feed hole corresponding to the cavity containing the volatile substance is provided with a sealing soft plug.

[0008] Furthermore, the fluid channel is divided into a first layer fluid channel and a second layer fluid channel, and a plurality of first micro-channels are provided on the lower end surface of the box body, and the first micro-channels are connected to the cavity through vias;

[0009] The box body also includes a lower cover plate covering the bottom of the box body, the lower end surface of the lower cover plate is provided with a plurality of second micro-channels, the lower end surface of the lower cover plate is covered with a bottom sealing film for sealing, the second micro-channels and the bottom sealing film cooperate to form a second layer of fluid channels, and the first micro-channels and the upper end surface of the lower cover plate cooperate to form a first layer of fluid channels.

[0010] Furthermore, an amplification detection area is also provided on the lower cover plate, and the amplification detection area includes an amplification detection array and a detection microchannel connected to the amplification detection array, and the upper end surface of the amplification detection area is covered with a detection area sealing film for sealing.

[0011] Preferably, the second microchannel is provided with a plurality of fluid state cavities, and the box body is provided with fluid observation windows corresponding to the fluid state cavities.

[0012] Furthermore, the nucleic acid detection card box also includes a rotating mechanism, and the rotating mechanism controls the fluid channels to be selectively connected by rotation.

[0013] Specifically, the rotating mechanism includes a soft pad and a rotating valve connected in sequence, the lower end surface of the soft pad is provided with a plurality of positioning holes, the positioning holes and a plurality of positioning columns provided on the box body cooperate to fix the rotating mechanism, the soft pad is connected to the fluid channel through a plurality of through holes, respectively, the lower end surface of the rotating valve is provided with a liquid flow channel and a gas flow channel, the liquid flow channel can be selectively connected to the fluid channel through the through holes, and the gas flow channel can be selectively connected to the fluid channel through the through holes.

[0014] Furthermore, the rotation mechanism also includes a limit ring and a limit wall. The lower end surface of the limit ring is set as a keying area and a smooth area. The smooth area presses the edge of the rotary valve for sealing. The keying area is connected to the limit wall. The limit ring and the limit wall cooperate to form a rotation space.

[0015] Furthermore, a gas inlet and outlet channel is provided on the box body, and the gas inlet and outlet channel can be selectively connected to the gas flow channel or cavity. The lower port of the gas inlet and outlet channel is covered with a waterproof breathable membrane. The gas inlet and outlet channel controls the flow direction of the substance used for nucleic acid extraction by providing gas power.

[0016] Furthermore, the box body has a length ranging from 50 mm to 70 mm, and / or a width ranging from 20 mm to 40 mm, and / or a height ranging from 15 mm to 30 mm.

[0017] The present invention also provides an assembly process for a nucleic acid detection cartridge, comprising the following steps:

[0018] Step 1: The lower cover plate is bonded to the box body to form a fluid channel;

[0019] Step 2: The box body and the upper cover are bonded to form a complete cavity;

[0020] Step 3: Assemble the rotating mechanism, install the cushion into the limiting wall, fix the rotating mechanism, ensure that the through holes on the cushion correspond to the through holes at the corresponding positions on the upper end surface of the box body, and then install the rotating valve above the cushion;

[0021] Step 4: adding liquid, adding the substances used for nucleic acid extraction into the cavity one by one, and then sealing;

[0022] Step 5: Add primers to the amplification detection array and seal it.

[0023] Furthermore, the assembly process of step one includes:

[0024] First, the bottom sealing film is bonded to the lower cover plate to form a second layer of fluid channels;

[0025] Secondly, the waterproof and breathable membrane is covered to the gas inlet and outlet channel position at the lower end surface of the box body;

[0026] Again, the lower cover plate is bonded to the box body to form a first layer of fluid channels.

[0027] The present invention realizes the molecular diagnosis POCT of "sample in, result out" in the true sense. It adopts the design of a fully enclosed cartridge, with a sophisticated structure and greatly reduced costs. All reagents used are pre-installed in the test cartridge and do not need to be added separately. It realizes the integration of extraction + amplification + detection, and at the same time has low requirements for supporting instruments, and only simple operations are required to realize the full process of molecular diagnosis. The present invention also adopts multiple PCR technology, reagent room temperature technology, and rapid extraction and purification technology. It can realize the full process of nucleic acid extraction and detection without the need for a professional molecular biology laboratory. It has a wide range of uses and is suitable for various scenarios, from the emergency department of a tertiary hospital to the community and township health center, and is particularly suitable for environments where conventional instruments are not suitable for work. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 It is a schematic diagram of the overall structure of a nucleic acid detection cartridge of the present invention;

[0029] Figure 2 This is an exploded view from above of a nucleic acid detection cartridge of the present invention;

[0030] Figure 3 This is an exploded view from the bottom of a nucleic acid detection cartridge of the present invention;

[0031] Figure 4 It is a front structural schematic diagram of the box body of the present invention;

[0032] Figure 5 It is a schematic diagram of the reverse structure of the box body of the present invention;

[0033] Figure 6 It is a bottom view of the box body of the present invention;

[0034] Figure 7 It is a top view of the box body of the present invention;

[0035] Figure 8 It is a front structural schematic diagram of the upper cover plate of the present invention;

[0036] Fig. 9 It is a schematic diagram of the reverse structure of the upper cover plate of the present invention;

[0037] Fig.10 It is a front structural schematic diagram of the lower cover plate of the present invention;

[0038] Fig.11 It is a schematic diagram of the reverse structure of the lower cover plate of the present invention;

[0039] Fig.12 It is a front structural schematic diagram of the rotary valve of the present invention;

[0040] Fig.13 It is a schematic diagram of the reverse structure of the rotary valve of the present invention;

[0041] Fig.14 It is a front structural schematic diagram of the soft cushion of the present invention;

[0042] Fig.15 It is a schematic diagram of the reverse structure of the cushion of the present invention;

[0043] Fig.16 It is a front structural schematic diagram of the limiting ring of the present invention;

[0044] Fig.17 It is a schematic diagram of the reverse structure of the limiting ring of the present invention;

[0045] Fig.18 is a schematic diagram of the use of the rotary valve of the present invention in the initial position;

[0046] Fig.19 is a schematic diagram of the use of the rotary valve of the present invention in step 2 of the use method;

[0047] In the figure, 1 is a sampling tube; 2 is a box body; 3 is a rotating mechanism; 21 is an upper cover plate; 22 is a box body; 23 is a lower cover plate; 221 is a first microchannel; 211 is a feed hole; 212 is a top sealing film; 213 is a sealing plug; 214 is a reagent chamber cover; 121 is an amplification detection array; 122 is a detection microchannel; 123 is a detection area sealing film; 231 is a second microchannel; 232 is a bottom sealing film; 2311 is a fluid state chamber; 2312 is a fluid observation window; 31 is a limit ring; 32 is a rotating valve; 33 is a soft plug; Pad; 34 limiting wall; 311 bonding area; 312 smooth area; 321 long groove; 322 short groove; 331 positioning hole; 332 positioning column; 101 magnetic bead chamber; 102 extraction chamber; 103 waste liquid chamber; 104 sample chamber; 105 cleaning chamber 1; 106 cleaning chamber 2; 107 elution chamber; 108 reagent chamber; 109 spare chamber; 1010 air hole A; 1011 air hole B; 1013 air hole C; 1012 mounting hole; 1014 waterproof and breathable membrane. DETAILED DESCRIPTION

[0048] The following is a description of the implementation of the present invention by specific embodiments. A person skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by a person skilled in the art without creative work are within the scope of protection of the present invention.

[0049] To facilitate those skilled in the art to understand the present invention, specific implementations of the present invention are described below with reference to the accompanying drawings.

[0050] See also Figures 1 to 19 , discloses a nucleic acid detection card box, including a sampling tube 1 and a box body 2, wherein the box body 2 is provided with a plurality of cavities for containing substances used for nucleic acid extraction and a plurality of fluid channels connected to the cavities, the sampling tube 1 is connected to one of the cavities, and the plurality of fluid channels can be selectively connected.

[0051] The nucleic acid detection card box of the present invention is very delicate in design, flexible in use, and greatly reduces various costs. Figure 1 As shown, the length range (L) of the box body 2 is 50mm-70mm, preferably 70mm; the width range (W) is 20mm-40mm, preferably 33mm; and the height range (H) is 15mm-30mm, preferably 20mm.

[0052] Specifically, the box body 2 is an integral structure that is roughly square, and is composed of a number of internal cavities, microchannels, through holes, etc., and is provided with at least a magnetic bead chamber 101, an extraction chamber 102, a waste liquid chamber 103, a sample chamber 104, a cleaning chamber 105, a cleaning chamber 2 106, an elution chamber 107, a reagent chamber 108, a spare chamber 109, and an installation hole 1012, wherein the sampling tube 1 is connected to the sample chamber 104.

[0053] like Figure 2 and Figure 3 As shown, the box body 2 includes a box body 22 and an upper cover plate 21 covering the box body 22. The above-described plurality of cavities are arranged inside the box body 22. The lower end surface (reverse surface) of the box body 22 is provided with a plurality of first micro-channels 221 of different sizes. The first micro-channels 221 are sequentially connected with the corresponding cavities through a plurality of vias, so that each group of cavities and the corresponding first micro-channels 221 are independent of each other and do not interfere with each other, forming an independent channel.

[0054] Specifically, the upper cover plate 21 is a flat plate, provided with a plurality of feed holes 211 corresponding to the cavities, and the upper end surface of the upper cover plate 21 is covered with a top sealing film 212 for sealing. In a preferred embodiment, the feed holes 211 corresponding to the cavities containing volatile liquids are provided with sealing plugs 213 before being covered with the top sealing film 212 to prevent the liquid from volatilizing, such as the magnetic bead chamber 101, the first cleaning chamber 105, and the second cleaning chamber 106.

[0055] Furthermore, the box body 2 also includes a lower cover plate 23 covering the bottom of the box body 22. The lower cover plate 23 is a flat plate, divided into two sides, the upper side is the front side, and is provided with an amplification detection area, including an amplification detection array 121 (through the upper and lower sides), and a detection micro-channel 122 connected to the amplification detection array. The upper end surface of the amplification detection area is covered with a detection area sealing film 123 for sealing. The lower side is the back side, and is provided with a plurality of second micro-channels 231 of different sizes, wherein the second micro-channels 231 are connected through a plurality of vias on the front and back sides, and connected to the detection micro-channels 122, so that each group of second micro-channels 231 are independent of each other, do not interfere with each other, and form an independent channel. The lower end surface (reverse side) of the lower cover plate 23 is covered with a bottom sealing film 232 for sealing the second micro-channels 231 and the amplification detection area.

[0056] The fluid channel is divided into a first layer fluid channel and a second layer fluid channel. The second microchannel 231 and the bottom sealing film 232 cooperate to form the second layer fluid channel, and the first microchannel 221 and the upper end surface (front face) of the lower cover plate 23 cooperate to form the first layer fluid channel. The first layer fluid channel and the second layer fluid channel cooperate to form a channel that can conduct between the cavities. In one embodiment, the second microchannel 231 is provided with a plurality of fluid state cavities 2311, and the box body 22 is provided with a fluid observation window 2312 corresponding to the fluid state cavity 2311, so as to dynamically observe the fluid state during the reaction process.

[0057] Furthermore, the nucleic acid detection cartridge also includes a rotating mechanism 3, which is composed of a limiting ring 31, a rotating valve 32, a cushion 33, and a limiting wall 34. The limiting ring 31 and the limiting wall 34 enclose a rotating space.

[0058] For more information, see Fig.14 and Fig.15 The soft pad 33 is made of a smooth and elastic material, such as styrene, styrene-isoprene-styrene block copolymer, polypropylene, thermoplastic polyurethane and other elastomeric materials. The lower end surface is provided with a plurality of positioning holes 331, and the positioning holes 331 and a plurality of positioning posts 332 provided on the box body 22 cooperate to fix the rotating mechanism 3. The soft pad 33 is also provided with a plurality of through holes that penetrate up and down, and the positions of the through holes correspond to the through holes at the corresponding positions of the box body 22 one by one to achieve penetration.

[0059] For more information, see Fig.12 and Fig.13 The rotary valve 32 is an integral part and is the only moving part in the rotary mechanism 3. The lower end surface of the rotary valve 32 is provided with two long grooves 321 and short grooves 322 of different lengths. The rotary valve 32 can rotate so that the long groove 321 cooperates with the through hole on the soft pad 33 to form a liquid flow path, and the liquid flow path can be selectively connected to each cavity; similarly, the rotary valve 32 can also rotate so that the short groove 322 cooperates with the through hole on the soft pad 33 to form a gas flow path, and the gas flow path can also be selectively connected to each cavity.

[0060] For more information, see Fig.16 and Fig.17 The lower end surface of the limiting ring 31 is set as a bonding area 311 and a smooth area 312. The bonding area 311 and the upper surface of the limiting wall 34 are completely fixed using a bonding process. The smooth area 312 will press down the rotating valve 32, forcing the lower surface of the rotating valve 32 to press down the soft pad 33 to deform, forming a certain pre-tightening force to play a sealing role.

[0061] Furthermore, the box body 22 is also provided with a gas inlet and outlet channel, including air hole A1010, air hole B1011 and waste liquid chamber outlet hole C1013, air hole A1010 and air hole B1011 penetrate the upper cover plate 21 and the top sealing film 212, and the waste liquid chamber outlet hole C1013 is connected to the waste liquid chamber. By rotating the rotary valve 32, the air hole A1010 or the air hole B1011 can be connected to the gas flow channel, and the air hole A1010 or the air hole B1011 can also be connected to the extraction chamber or the reagent chamber; the air hole A1010, the air hole B1011 and the waste liquid chamber outlet hole C1013 cooperate to inlet and outlet air, thereby providing gas power to control the flow direction of the substances used for nucleic acid extraction in each cavity. The lower port of the gas inlet and outlet channel, i.e., the corresponding lower end surface of the box body 22, is covered with a waterproof breathable membrane 1014. The waterproof breathable membrane 1014 is used to isolate the internal and external environments of the test card box, and has the function of blocking liquid and allowing air to pass through, thereby ensuring that liquids, aerosols and other organic or harmful substances inside the test card box and interfering substances in the external environment of the test card box cannot enter or exit, thereby avoiding pollution and interference.

[0062] The present invention also discloses an assembly process of a nucleic acid detection cartridge, comprising the following steps:

[0063] Step 1: The bottom sealing film 232 is bonded to the lower cover plate 23 to form a second layer of fluid channels;

[0064] Step 2: Cover the waterproof breathable membrane 1014 to the corresponding gas inlet and outlet channel position on the lower end surface of the box body 22;

[0065] Step 3: The lower cover plate 23 is bonded to the lower end surface of the box body 22 to form a first layer of fluid channel;

[0066] Step 4: The box body 22 and the upper cover plate 21 are bonded to form a complete cavity;

[0067] Step 5: Assemble the rotating mechanism 3:

[0068] First, the cushion 33 is installed into the limiting wall 34, and then the positioning column 332 on the box body 22 cooperates with the positioning hole 331 on the cushion 33 to connect and fix the position, and close to the bottom surface, to ensure that the through hole on the cushion 33 corresponds to the through hole in the limiting wall 34 and the upper end surface of the box body 22 one by one; then, the rotary valve 32 and the limiting ring 31 are installed in sequence, and the bonding area of ​​the lower end surface of the limiting ring 31 and the upper end surface of the limiting wall 34 are completely fixed by a bonding process, and at the same time, the smooth area 312 of the lower end surface of the limiting ring 31 presses down the rotary valve 32, forcing the lower surface of the rotary valve 32 to press down the cushion 33 to deform, forming a certain pre-tightening force, which plays a sealing role;

[0069] Step 6: Add liquid, add the substances used for nucleic acid extraction one by one through the feed hole 211 of the upper cover plate 21, seal the feed hole 211 with a sealing plug 213 for volatile liquid, and then use the top sealing film 212 to bond and seal with the upper cover plate 21;

[0070] Step 7: Add reagents to the reagent chamber and seal it with the reagent chamber cover 214;

[0071] Step eight: Add primers to the amplification detection array and use the detection area sealing film 123 to bond and seal.

[0072] The bonding described in the above steps refers to fixing two identical or different materials together into one, such as pressure-sensitive adhesive, photosensitive adhesive, heat-sensitive adhesive, double-sided tape, thermal bonding, ultrasonic bonding, laser bonding, protrusion coupling to groove, etc.

[0073] The hard material and film material used in the detection card box can be made of high molecular polymer materials such as polypropylene, cycloolefin copolymer, cycloolefin polymer, polymethyl methacrylate, polystyrene or polycarbonate.

[0074] For ease of understanding, the method of using the nucleic acid detection cartridge in the present invention is as follows:

[0075] Step 1: Rotate the rotary valve 32, the extraction chamber and the magnetic bead chamber are connected through the fluid channel, the air hole A1010 exhausts air, the air hole B1011 intakes air, the liquid in the magnetic bead chamber enters the extraction chamber, and after mixing, the external magnet adsorbs the magnetic beads;

[0076] Step 2: Rotate the rotary valve 32, the extraction chamber and the waste liquid chamber are connected through the fluid channel, the air hole A1010 takes in air, the waste liquid chamber air hole C1013 discharges air, and the waste liquid in the extraction chamber is discharged into the waste liquid chamber;

[0077] Step 3: Rotate the rotary valve 32, the extraction chamber and the sample chamber are connected through the fluid channel, the air hole A1010 exhausts air, and the air hole B1011 intakes air, the sample in the sample chamber enters the extraction chamber, the sample is mixed with the magnetic beads, and after the magnetic beads absorb the nucleic acid in the sample, the external magnet absorbs the magnetic beads;

[0078] Step 4: Repeat step 2;

[0079] Step 5: Rotate the rotary valve 32, the extraction chamber and the cleaning chamber 1 are connected through the fluid channel, the air hole A1010 exhausts air, the air hole B1011 intakes air, the cleaning liquid in the cleaning chamber 1 enters the extraction chamber, the nucleic acid adsorbed by the magnetic beads is cleaned, and the external magnet adsorbs the magnetic beads;

[0080] Step 6: Repeat step 2;

[0081] Step 7: Rotate the rotary valve 32, the extraction chamber and the second cleaning chamber are connected through the fluid channel, the air hole A1010 exhausts air, the air hole B1011 intakes air, the cleaning liquid in the second cleaning chamber enters the extraction chamber, the nucleic acid adsorbed by the magnetic beads is cleaned, and the external magnet adsorbs the magnetic beads;

[0082] Step 8: Repeat step 2;

[0083] Step nine: Rotate the rotary valve 32, the extraction chamber and the elution chamber are connected through the fluid channel, the air hole A1010 exhausts air, the air hole B1011 intakes air, the eluent in the elution chamber enters the extraction chamber, the eluent is mixed with the magnetic beads, the target nucleic acid substance adsorbed by the magnetic beads is eluted, and the external magnet adsorbs the magnetic beads;

[0084] Step 10: Rotate the rotary valve 32, the extraction chamber and the reagent chamber are connected through the fluid channel, the air hole A1010 takes in air, the air hole B1011 lets out air, the eluent mixed with the target nucleic acid substance enters the reagent chamber, and mixes with the freeze-dried reagent in the reagent chamber; after the mixing is completed, the air hole A1010 lets out air, the air hole B1011 lets in air, the external magnet remains in the adsorption state, and the mixed reagent in the reagent chamber enters the extraction chamber;

[0085] Step 11: Rotate the rotary valve 32, the extraction chamber and the amplification detection array are connected through the fluid channel, the air hole A1010 takes in air, the air hole B1011 lets out air, and the mixed reagent solution in the extraction chamber enters the amplification detection array;

[0086] Step 12: Rotate the rotary valve 32 to completely seal the amplification and detection area, and then start nucleic acid amplification and detection.

[0087] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made on the basis of the present invention. Therefore, these modifications or improvements made on the basis of not departing from the spirit of the present invention all belong to the scope of protection claimed by the present invention. The above-described embodiments of the present invention do not constitute a limitation on the scope of protection of the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the scope of protection of the claims of the present invention.

Claims

1. A nucleic acid detection cartridge, characterized in that: The invention comprises a sampling tube (1) and a box body (2), wherein the box body (2) is provided with a plurality of cavities for containing substances used for nucleic acid extraction and a plurality of fluid channels connected to the cavities, wherein the sampling tube (1) is connected to one of the cavities, and the fluid channels can be selectively connected; the cavities comprise a magnetic bead chamber (101), an extraction chamber (102), a waste liquid chamber (103), a sample chamber (104), a first cleaning chamber (105), a second cleaning chamber (106), an elution chamber (107), a reagent chamber (108), and a standby chamber (109), wherein the sampling tube (1) is connected to the sample chamber (104), The nucleic acid detection cartridge further comprises a rotating mechanism (3), wherein the rotating mechanism (3) controls the fluid channels to selectively communicate with each other through rotation; The rotating mechanism (3) comprises a soft pad (33) and a rotating valve (32) connected in sequence. The lower end surface of the soft pad (33) is provided with a plurality of positioning holes (331). The positioning holes (331) and a plurality of positioning posts (332) provided on the box body (2) cooperate to fix the rotating mechanism (3). The soft pad (33) is connected to the fluid channel through a plurality of through holes. The lower end surface of the rotating valve (32) is provided with a long groove (321) and a short groove (322). The rotating valve (32) is configured to rotate so that the long groove (321) cooperates with the through hole on the soft pad (33) to form a liquid flow channel, and the short groove (322) cooperates with the through hole on the soft pad (33) to form a gas flow channel. The liquid flow channel can selectively communicate with the fluid channel, and the gas flow channel can selectively communicate with the fluid channel. The box body (2) comprises a box body (22) and an upper cover plate (21) covering the box body (22), the upper cover plate (21) being provided with a feed hole (211) corresponding to the cavity, the fluid channel being divided into a first layer fluid channel and a second layer fluid channel, a plurality of first micro-channels (221) being provided on the lower end surface of the box body (22), the first micro-channels (221) being connected to the cavity through vias; The box body (2) further comprises a lower cover plate (23) covering the lower part of the box body (22); a plurality of second microchannels (231) are arranged on the lower end surface of the lower cover plate (23); a bottom sealing film (232) is covered on the lower end surface of the lower cover plate (23) for sealing; the second microchannels (231) and the bottom sealing film (232) cooperate to form a second layer of fluid channels; the first microchannels (221) and the upper end surface of the lower cover plate (23) cooperate to form a first layer of fluid channels; the first layer of fluid channels The body channel and the second layer fluid channel cooperate to form a channel that can conduct between the cavities; the front side of the lower cover plate (23) is also provided with an amplification detection area, the amplification detection area includes an amplification detection array (121) and a detection microchannel (122) connected to the amplification detection array (121), and the upper end of the amplification detection area is covered with a detection area sealing film (123) for sealing; the front and back sides of the lower cover plate (23) are connected to the second microchannel (231) through a plurality of through holes and connected to the detection microchannel (122); The rotating mechanism (3) further comprises a limiting ring (31) and a limiting wall (34); the lower end surface of the limiting ring (31) is arranged as a keying area (311) and a smooth area (312); the smooth area (312) is pressed against the edge of the rotating valve (32) for sealing; the keying area (311) is connected to the limiting wall (34); the limiting ring (31) and the limiting wall (34) cooperate to form a rotating space; The box body (2) is also provided with a plurality of gas inlet and outlet channels, including an air hole A (1010), an air hole B (1011) and a waste liquid chamber air outlet C (1013). The rotary valve (32) is rotated so that the gas inlet and outlet channels can be selectively connected to the gas flow channel or the cavity. The lower side port of the gas inlet and outlet channels is covered with a waterproof and breathable membrane (1014). The gas inlet and outlet channels control the flow direction of the substances used for nucleic acid extraction by providing gas power. The method for using the nucleic acid detection cartridge is as follows: Step 1: Rotate the rotary valve (32), the extraction chamber and the magnetic bead chamber are connected through the fluid channel, the air hole A (1010) exhausts air, the air hole B (1011) intakes air, the liquid in the magnetic bead chamber enters the extraction chamber, and after mixing, the external magnet adsorbs the magnetic beads; Step 2: Rotate the rotary valve (32), so that the extraction chamber and the waste liquid chamber are connected through the fluid channel, air enters through the air hole A (1010), and air is discharged through the waste liquid chamber air outlet C (1013), and the waste liquid in the extraction chamber is discharged into the waste liquid chamber; Step 3: Rotate the rotary valve (32), the extraction chamber and the sample chamber are connected through the fluid channel, the air is discharged from the air hole A (1010) and the air is taken in from the air hole B (1011), the sample in the sample chamber enters the extraction chamber, the sample is mixed with the magnetic beads, and after the magnetic beads absorb the nucleic acid in the sample, the external magnet absorbs the magnetic beads; Step 4: Repeat step 2; Step 5: Rotate the rotary valve (32), the extraction chamber and the cleaning chamber 1 are connected through the fluid channel, the air is discharged from the air hole A (1010) and the air is taken in from the air hole B (1011), the cleaning liquid in the cleaning chamber 1 enters the extraction chamber, the nucleic acid adsorbed by the magnetic beads is cleaned, and the external magnet adsorbs the magnetic beads; Step 6: Repeat step 2; Step 7: Rotate the rotary valve (32), the extraction chamber and the second cleaning chamber are connected through the fluid channel, the air is discharged from the air hole A (1010) and the air is taken in from the air hole B (1011), the cleaning liquid in the second cleaning chamber enters the extraction chamber, the nucleic acid adsorbed by the magnetic beads is cleaned, and the external magnet adsorbs the magnetic beads; Step 8: Repeat step 2; Step nine: rotating the rotary valve (32), the extraction chamber and the elution chamber are connected through the fluid channel, the air hole A (1010) exhausts air, the air hole B (1011) intakes air, the eluent in the elution chamber enters the extraction chamber, the eluent is mixed with the magnetic beads, the target nucleic acid substance adsorbed by the magnetic beads is eluted, and the external magnet adsorbs the magnetic beads; Step 10: Rotate the rotary valve (32), the extraction chamber and the reagent chamber are connected through the fluid channel, the pore A (1010) takes in air, the pore B (1011) lets out air, the eluent mixed with the target nucleic acid substance enters the reagent chamber, and mixes with the freeze-dried reagent in the reagent chamber; after the mixing is completed, the pore A (1010) lets out air, the pore B (1011) lets in air, the external magnet remains in the adsorption state, and the mixed reagent in the reagent chamber enters the extraction chamber; Step 11: Rotate the rotary valve (32), the extraction chamber and the amplification detection array are connected through the fluid channel, the air hole A (1010) takes in air, the air hole B (1011) lets out air, and the mixed reagent solution in the extraction chamber enters the amplification detection array; Step 12: Rotate the rotary valve (32) to completely seal the amplification and detection area, and then start nucleic acid amplification and detection.

2. A nucleic acid detection cartridge according to claim 1, characterized in that: The upper end surface of the upper cover plate (21) is covered with a top sealing film (212) for sealing.

3. A nucleic acid detection cartridge according to claim 2, characterized in that: The feeding hole (211) corresponding to the cavity containing the volatile substance is provided with a sealing soft plug (213).

4. A nucleic acid detection cartridge according to claim 1, characterized in that: The second microchannel (231) is provided with a plurality of fluid state chambers (2311), and the box body (22) is provided with fluid observation windows (2312) corresponding to the fluid state chambers (2311).

5. A nucleic acid detection cartridge according to claim 1, characterized in that: The box body (2) has a length ranging from 50 mm to 70 mm, and / or a width ranging from 20 mm to 40 mm, and / or a height ranging from 15 mm to 30 mm.

6. An assembly process for the nucleic acid detection cartridge according to any one of claims 1 to 5, characterized in that: The assembly process comprises the following steps: Step 1: The lower cover plate (23) is bonded to the box body (22) to form a fluid channel; Step 2: The box body (22) and the upper cover plate (21) are bonded to form a complete cavity; Step 3: Assemble the rotating mechanism (3), install the soft pad (33) into the limiting wall (34) to fix the rotating mechanism (3), ensure that the through holes on the soft pad (33) correspond to the through holes at the corresponding positions on the upper end surface of the box body (22), and then install the rotating valve (32) above the soft pad (33); Step 4: adding liquid, adding the substances used for nucleic acid extraction into the cavity one by one, and then sealing; Step 5: Add primers to the amplification detection array and seal it.

7. The assembly process of a nucleic acid detection cartridge according to claim 6, characterized in that: The assembly process of step 1 includes: First, the bottom sealing film (232) is bonded to the lower cover plate (23) to form a second layer of fluid channels; Secondly, the waterproof and breathable membrane (1014) is covered to the gas inlet and outlet channel position on the lower end surface of the box body (22); Again, the lower cover plate (23) is bonded to the box body (22) to form a first layer of fluid channels.

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