Nucleic acid detection kit and manufacturing process thereof

Abstract

A nucleic acid detection kit comprising a lower circular fixed lid (1), a circular kit (2) inserted inside the lower circular fixed lid (1), a sealing membrane (3) provided on the top of the circular kit (2), a lower latex plastic pressing lid (4) provided on the lower end of the sealing membrane (3), a bottom of the lower latex plastic pressing lid (4) in pressing contact with the lower end of the sealing membrane (3) and the bottom of the sealing membrane (3) in pressing contact with the top of the circular kit (2), a first bolt (5) rotatably connected at an equal distance to the outside of the lower circular fixed lid (1) by a bearing, and the first bolt (5) is screwed into the lower latex plastic pressing lid (4) by a first screw hole. The nucleic acid detection kit can greatly improve the sealing efficiency, is convenient to operate, and can detect gas leakage after sealing is completed. Compared with conventional detection kits, the sealing quality is greatly improved and the sealing efficiency is improved. During processes such as nucleic acid extraction, amplification reaction, and fluorescence detection, the sealing film can prevent aerosols from leaking from the kit, thereby preventing cross-contamination.

Description

[Technical field]

[0001] The present invention relates to the technical field of nucleic acid detection kits, and in particular to a nucleic acid detection kit and a manufacturing process thereof. [Background technology]

[0002] Nucleic acids are the carriers of genetic information in organisms and are essential components of all known life forms. DNA nucleic acids and RNA nucleic acids are mainly present in the cell nucleus and exist in a state bound to proteins. With the rapid development of molecular biology, the research and analysis of nucleic acids has been continuously promoted and applied in fields such as clinical diagnosis, food safety, environmental detection, agriculture, forestry, and animal husbandry. Polymerase chain reaction (PCR) technology is a technology for rapid amplification of DNA in vitro. This technology can be applied to basic research such as gene isolation, cloning, and nucleic acid sequence analysis, as well as any field related to DNA or RNA, such as disease diagnosis.

[0003] Nucleic acid analysis technology, as a powerful molecular diagnostic technology, has important applications in pathogen diagnosis, infectious disease prevention, disease monitoring and treatment, etc. Compared with ordinary immunological diagnostic methods, nucleic acid analysis has excellent advantages such as high specificity and high sensitivity. Most target genes have low concentrations and are difficult to directly analyze and detect, so nucleic acid analysis methods such as PCR and LAMP based on the principle of nucleic acid amplification have become the main means of current nucleic acid analysis. The complete process generally includes three main links: nucleic acid extraction, nucleic acid amplification and nucleic acid detection. In the existing nucleic acid detection kit, it is difficult to integrate the entire process of nucleic acid extraction and nucleic acid detection in the same closed space. As a result, in the process of nucleic acid detection, the entire process of nucleic acid extraction and nucleic acid detection needs to be separated, which will cause cross-contamination of the sample aerosol, which is prone to false positives in the nucleic acid detection of the subsequent sample, and the detection efficiency is low. Therefore, in order to solve the above technical problems, it is necessary to provide a nucleic acid detection kit and its manufacturing process. Summary of the Invention

[0004] To solve the above technical problems, the present invention provides a nucleic acid detection kit and a manufacturing process thereof.

[0005] The nucleic acid detection kit provided by the present invention includes a lower circular fixed cover, a circular kit is inserted inside the lower circular fixed cover, a sealing film is provided on the top of the circular kit, a lower latex plastic pressing cover is provided on the lower end of the sealing film, the bottom of the lower latex plastic pressing cover is in pressing contact with the lower end of the sealing film, and the bottom of the sealing film is in pressing contact with the top of the circular kit, and a first bolt is rotatable at an equal distance by a bearing on the outside of the lower circular fixed cover. The first bolt is screwed into the lower latex plastic pressing lid through the first screw hole, a metal rod receiving hole is provided in the center of the circular kit, an upper lid is provided above the sealing film, an injection port is provided on one side of the circular kit, a rubber plug is tightly connected to the center of the injection port, a metal rod is fixed to the center of the upper lid, the lower end of the metal rod is inserted into the metal rod receiving hole, and an upper circular fixing lid is fixed to the outer wall of the upper lid, a second bolt is rotatably connected to an outer ring of the upper ring fixing cover by a bearing at an equal distance, and the second bolt is screwed into the upper latex plastic pressing cover by a second screw hole; an embedded tube mounting hole is opened on one side of the circular kit, a PCR tube is fixed to the outside of the embedded tube mounting hole at the bottom of the circular kit, and an embedded tube assembly is provided on the inside of the embedded tube mounting hole; shallow tubes are arranged at equal intervals on the side of the circular kit far from the PCR tube; and a U-shaped aluminum foil film is provided on the side of the circular kit close to the shallow tube; a detection hole is opened on one side of the upper cover, and a detection plug is tightly fitted and connected to the inner wall of the detection hole.

[0006] Preferably, a first connection mechanism is fixed to the bottom of the lower ring fixed lid, and the first connection mechanism is used to drive the simultaneous rotation of a first bolt, and a second connection mechanism is provided on the top of the upper ring fixed lid, and the second connection mechanism is used to drive the simultaneous rotation of a second bolt.

[0007] Preferably, the embedded tube assembly includes an embedded tube body, an aluminum foil film B, and an aluminum foil film A, the embedded tube body is provided on the inner wall of the embedded tube mounting hole, the aluminum foil film B is heat sealed to the bottom of the embedded tube body, the aluminum foil film A is heat sealed to the top of the embedded tube body, and the bottom of the aluminum foil film B is in pressure contact with the top of a PCR tube.

[0008] Preferably, the first connection mechanism includes a bottom connecting plate, an internal gear ring, a first connecting shaft, a first gear, a worm wheel, a support block, a worm, and a first rotating wheel, wherein a bottom connecting plate is fixed to the bottom of the lower ring fixing cover, an internal gear ring is rotatably connected to the lower surface of the bottom connecting plate by a bearing, a first connecting shaft is fixed to the middle part of the lower surface of the first bolt, a first gear is fixed to the middle part of the first connecting shaft, all six of the first gears are meshed and connected to the internal gear ring, a worm wheel is fixed to the bottom of the internal gear ring, support blocks are fixed symmetrically to one side of the lower surface of the bottom connecting plate, a worm is rotatably connected between the two support blocks by a bearing, the worm is meshed and connected to the worm wheel, and one end of the worm passes through the support block and is fixed to the first rotating wheel.

[0009] Preferably, first positioning pillars are fixed at equal intervals to the top of the lower ring fixing cover, and first positioning sleeves are fixed at equal intervals to the outer wall of the lower latex plastic pressing cover, and the first positioning pillars are inserted into the first positioning sleeves.

[0010] Preferably, the second connection mechanism includes a second connecting shaft, a second gear, a rotating ring, a gear ring, and a second rotating wheel, a second connecting shaft is fixed to the middle of the upper surface of the second bolt, a second gear is fixed to the upper end of the second connecting shaft, a rotating ring is rotatably connected to the middle of the upper surface of the upper ring fixing cover by a bearing, a gear ring is fixed to the outer wall of the upper end of the rotating ring, and all six second gears are meshed and connected to the gear ring, and a second rotating wheel is fixed to the top of one second connecting shaft on one side of the upper ring fixing cover.

[0011] Preferably, second positioning pillars are fixed at equal intervals to the outer wall of the upper circular fixed cover, second positioning sleeves are fixed at equal intervals to the outer wall of the upper latex plastic pressing cover, the second positioning pillars are inserted into the second positioning sleeves, and the first rotating wheel and the second rotating wheel are both plum-shaped.

[0012] Preferably, both the top of the first positioning post and the bottom of the second positioning post are provided with a chamfer.

[0013] Preferably, support posts are fixed at equal intervals to the bottom of the bottom connecting plate, and rubber friction pads are fixed to the bottom of the support posts.

[0014] Preferably, the sealing membrane comprises an elastic membrane or a non-elastic membrane.

[0015] The present invention further provides a process for producing a nucleic acid detection kit, comprising the following steps 1) to 5): 1) The injection port is blocked with a rubber plug, and the nucleic acid extraction reagent and magnetic beads in the circular kit are sealed with a U-shaped aluminum foil membrane, and the nucleic acid extraction reagent and magnetic beads are placed in the corresponding storage ports in the circular kit, and the magnetic beads may be replaced with a magnetic bead solution; 2) adding a liquid or solid reagent into the PCR tube, and then pressing the embedded tube assembly into the PCR tube; the order of steps 1) and 2) can be reversed; 3) The sealing film is adhered to the top of the circular kit and the bottom of the top cover and fixed by crimping; 4) Connect to an external gas source, introduce gas through the detection hole in the top cover, and detect the tightness of the sealing film. 5) If no gas leak is detected, the detection hole in the top cover is blocked with the detection plug.

[0016] In a preferred embodiment of the present invention, a desiccant is fixed in the space within the sealing film, and the desiccant is fixed to an upper cover or a base at a position that does not affect the movement of the metal rod and the inclusion of the reagent. The PCR tube has an outer wall thickness of 0.1-1.5 mm and a light transmittance of 60% or more to enable optical detection.

[0017] The top surface of the U-shaped aluminum foil membrane does not react with the reagent contained therein, and a PP plastic film is attached to the bottom surface, allowing it to be stored while in contact with the liquid and solid in the shallow tube. The aluminum foil membrane A at the top of the buried tube body has one side in contact with the reagent, and the aluminum foil membrane B at the bottom can be in contact with the reagent on both sides. The order of sealing the buried tube assembly 16 body is to seal the aluminum foil membrane B at the bottom first, add the reagent to be embedded, and then seal the aluminum foil membrane A at the top. The liquid or solid reagent at the bottom is added to the PCR tube, and then the bottom surface of the aluminum foil membrane B is washed and the buried tube assembly is added, thereby realizing the sealing of the PCR tube.

[0018] Compared with the prior art, the nucleic acid detection kit and its manufacturing process provided in the present invention have the following beneficial effects:

[0019] The present invention provides a nucleic acid detection kit and a manufacturing process thereof.

[0020] 1. The nucleic acid detection kit provided by the present invention can greatly improve the sealing efficiency when sealing the nucleic acid detection reagent, is convenient to operate, and can detect gas leakage after sealing is completed, which greatly improves the sealing quality compared with the conventional detection kit.

[0021] 2. The nucleic acid detection kit provided by the present invention has a sealing membrane that prevents aerosols from leaking from the kit during the processes of nucleic acid extraction, amplification reaction, fluorescence detection, etc., thereby preventing cross-contamination.

[0022] 3. The nucleic acid detection kit provided in the present invention includes a complete process of nucleic acid extraction, nucleic acid amplification and nucleic acid detection, which can ensure the sealed environment of the nucleic acid detection reagent device, reduce the risk of sample cross-contamination, and effectively improve the detection efficiency of nucleic acid detection. [Brief description of the drawings]

[0023] [Figure 1] FIG. 1 is a schematic diagram of the overall structure provided in the present invention. [Diagram 2] FIG. 2 is a schematic diagram of the position structure of the upper latex plastic pressing cover provided by the present invention. [Diagram 3] 1 is a schematic diagram of the shape and structure of a sealing film provided by the present invention. [Figure 4] FIG. 2 is a schematic diagram of the position structure of the metal rod receiving hole provided in the present invention. [Diagram 5] FIG. 2 is a schematic diagram of the position structure of the embedded pipe mounting hole provided by the present invention. [Figure 6] FIG. 2 is a schematic diagram of the position structure of the PCR tube provided by the present invention. [Figure 7] FIG. 2 is a structural schematic diagram of the embedded tube assembly provided in the present invention. [Figure 8] FIG. 2 is a structural schematic diagram of a first connection mechanism provided in the present invention. [Figure 9] FIG. 2 is a structural schematic diagram of a second connection mechanism provided in the present invention. [Figure 10] FIG. 2 is an enlarged view of A provided by the present invention. [Figure 11] FIG. 2 is a schematic diagram of the shallow tube positioning structure provided in the present invention.

[0024] Explanation of symbols 1, lower circular ring fixed cover;2, circular kit;3, sealing membrane;4, lower latex plastic pressing cover;5, first bolt;6, metal rod receiving hole;7, upper cover;8, injection port;9, rubber stopper;10, metal rod;11, upper circular ring fixed cover;12, upper latex plastic pressing cover;13, second bolt;14, embedded tube mounting hole;15, PCR tube;16, embedded tube assembly;161, embedded tube body;162, aluminum foil film B;163, aluminum foil film A;17, shallow tube;18, U-shaped aluminum foil film;19, first connection mechanism;191, bottom connection plate;192, inner tooth ring;193, first connection shaft;1 94, first gear; 195, worm wheel; 196, support block; 197, worm; 198, first rotating wheel; 20, second connecting mechanism; 201, second connecting shaft; 202, second gear; 203, rotating ring; 204, gear ring; 205, second rotating wheel; 21, first positioning column; 22, first positioning sleeve; 23, second positioning column; 24, second positioning sleeve; 25, support column; 26, rubber friction pad; 27, detection hole; 28, detection plug; a, cleaning liquid storage port 1; b, cleaning liquid storage port 2; c, cleaning liquid storage port 3; d, cleaning liquid storage port 4; e, dissolving liquid storage tube. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] The present invention will be further described below with reference to the drawings and embodiments. In a specific implementation process, as shown in FIG. 1, FIG. 2, FIG. 3, FIG. 4, FIG. 5, FIG. 6 and FIG. 11, the nucleic acid detection kit includes a lower circular ring fixed cover 1. A circular kit 2 is inserted inside the lower circular ring fixed cover 1. A sealing film 3 is provided on the top of the circular kit 2, and a lower latex plastic pressing cover 4 is provided on the lower end of the sealing film 3. The bottom of the lower latex plastic pressing cover 4 is in pressing contact with the lower end of the sealing film 3, and the bottom of the sealing film 3 is in pressing contact with the top of the circular kit 2. A first bolt 5 is rotatably connected to the outside of the lower circular ring fixed cover 1 by a bearing at an equal distance. The first bolt 5 is screwed into the lower latex plastic pressing cover 4 through a first screw hole. A metal rod receiving hole 6 is provided in the middle of the circular kit 2. An upper cover 7 is provided above the sealing film 3. An injection port 8 is provided on one side of the circular kit 2. A rubber plug 9 is connected to the center of the injection port 8 by tight fit. A metal rod 10 is fixed to the center of the top cover 7. The lower end of the metal rod 10 is inserted into the metal rod receiving hole 6. The metal rod 10 may be inserted when the top cover 7 is injection molded, or may be fitted to the metal rod 10 after the top cover 7 is processed. An upper circular ring fixed cover 11 is fixed to the outer wall of the top cover 7, and an upper latex plastic pressing cover 12 is fixed to the upper end of the sealing film 3. The top of the upper latex plastic pressing cover 12 is in pressing contact with the upper end of the sealing film 3, and the top of the sealing film 3 is in pressing contact with the bottom of the top cover 7. In order to prevent the sealing film 3 from becoming thin and peeling off due to the pulling of the top cover 7, 3M double-sided tape is attached to the 45-degree (or other angle) surface of the sealing film 3 to increase friction. A second bolt 13 is rotatably connected to the outer ring of the upper circular ring fixed cover 11 by a bearing at an equal distance. The second bolt 13 is screwed into the upper latex plastic press cover 12 through a second screw hole. A buried tube mounting hole 14 is opened on one side of the circular kit 2. A PCR tube 15 is fixed to the outside of the buried tube mounting hole 14 at the bottom of the circular kit 2. A buried tube assembly 16 is provided inside the buried tube mounting hole 14. Shallow tubes 17 are arranged at equal intervals on the side of the circular kit 2 away from the PCR tubes 15. A U-shaped aluminum foil membrane 18 is provided on the side of the circular kit 2 close to the shallow tubes 17 at the top.The function of the U-shaped aluminum foil film 18 is to seal and store the contents, and to open a hole larger than the diameter of the metal bar 10 in the U-shaped aluminum foil film 18 by piercing the U-shaped aluminum foil film 18 with a probe when detecting using the kit. The U-shaped aluminum foil film 18 does not affect the vertical linear motion of the metal bar 10. The material of the sealed aluminum foil film may be other plastic films, etc. A first connection mechanism 19 is fixed to the bottom of the lower circular fixed cover 1. The first connection mechanism 19 is used to drive the simultaneous rotation of the first bolt 5. A second connection mechanism 20 is provided on the top of the upper circular fixed cover 11. The second connection mechanism 20 is used to drive the simultaneous rotation of the second bolt 13. A detection hole 27 is opened on one side of the upper cover 7. A detection plug 28 is connected to the inner wall of the detection hole 27 by tight fit.

[0026] Furthermore, according to the present invention, the efficiency of sealing can be greatly improved when sealing a nucleic acid detection reagent, making the operation convenient, and gas leakage can be detected after sealing is completed, thereby greatly improving the sealing quality compared to conventional detection kits.

[0027] 7, the embedded pipe assembly 16 includes an embedded pipe body 161, an aluminum foil film B 162, and an aluminum foil film A 163. The embedded pipe body 161 is provided on the inner wall of the embedded pipe mounting hole 14. The aluminum foil film B 162 is heat sealed to the bottom of the embedded pipe body 161. The aluminum foil film A 163 is heat sealed to the top of the embedded pipe body 161. The bottom of the aluminum foil film B 162 is in pressure contact with the top of the PCR tube 15.

[0028] The liquid or solid reagent in the PCR tube 15 is sealed by pressing the aluminum foil film B162 against a step on the PCR tube 15. A sealing material may be added between the step and the aluminum foil film B162. The embedded tube assembly 16 is fixed by pressing the circular kit 2 into the PCR tube 15, and then welding, gluing, structural fastening, or the like.

[0029] As shown in FIG. 8, the first connection mechanism 19 includes a bottom connection plate 191, an inner gear ring 192, a first connection shaft 193, a first gear 194, a worm wheel 195, a support block 196, a worm 197, and a first rotating wheel 198. The bottom connection plate 191 is fixed to the bottom of the lower ring fixing cover 1. The inner gear ring 192 is rotatably connected to the lower surface of the bottom connection plate 191 by a bearing. The first connection shaft 193 is fixed to the center of the lower surface of the first bolt 5. The first gear 194 is fixed to the center of the first connection shaft 193. All six of the first gears 194 are meshed and connected to the inner gear ring 192. The worm wheel 195 is fixed to the bottom of the inner gear ring 192. Support blocks 196 are symmetrically fixed to one side of the lower surface of the bottom connection plate 191. A worm 197 is rotatably connected by a bearing between the two support blocks 196. The worm 197 is meshed with a worm wheel 195. One end of the worm 197 passes through the support blocks 196 and is fixed to a first rotating wheel 198.

[0030] As shown in Fig. 1, first positioning posts 21 are fixed at equal intervals to the top of the lower ring fixed cover 1. First positioning sleeves 22 are fixed at equal intervals to the outer wall of the lower latex plastic pressing cover 4. The first positioning posts 21 are inserted into the first positioning sleeves 22 and are used to position the lower latex plastic pressing cover 4 when pressing the lower latex plastic pressing cover 4.

[0031] As shown in Figs. 9 and 10, the second connection mechanism 20 includes a second connecting shaft 201, a second gear 202, a rotating ring 203, a gear ring 204, and a second rotating wheel 205. The second connecting shaft 201 is fixed to the center of the upper surface of the second bolt 13. The second gear 202 is fixed to the upper end of the second connecting shaft 201. The rotating ring 203 is rotatably connected to the center of the upper surface of the upper ring fixing cover 11 by a bearing. The gear ring 204 is fixed to the outer wall of the upper end of the rotating ring 203. All six second gears 202 are meshed and connected to the gear ring 204. A second rotating wheel 205 is fixed to the top of one second connecting shaft 201 on one side of the upper ring fixing cover 11.

[0032] In addition, by providing the first connection mechanism 19 and the second connection mechanism 20, the first bolt 5 or the second bolt 13 can be rotated simultaneously by simply rotating the first rotating wheel 198 and the second rotating wheel 205 when pressing the sealing film 3, which greatly improves the sealing efficiency and is convenient to use.

[0033] In addition to the above-mentioned connection method, the pressing method of the latex plastic pressing lid may be such that the upper and lower circular fixed lids are changed to plastic ones and the upper and lower latex plastic pressing lids are welded to the upper and lower circular fixed lids, respectively, by ultrasonic welding or the like.

[0034] As shown in Fig. 1, second positioning posts 23 are fixed at equal intervals to the outer wall of the upper circular fixed cover 11. Second positioning sleeves 24 are fixed at equal intervals to the outer wall of the upper latex plastic pressing cover 12. The second positioning posts 23 are inserted into the second positioning sleeves 24 and are used to position the upper latex plastic pressing cover 12 when pressing the upper latex plastic pressing cover 12.

[0035] 8 and 10, the first rotating wheel 198 and the second rotating wheel 205 are both plum-shaped, which allows the first rotating wheel 198 or the second rotating wheel 205 to rotate easily.

[0036] 1, the top of the first positioning post 21 and the bottom of the second positioning post 23 are both chamfered, so that the first positioning post 21 can be easily inserted into the first positioning sleeve 22, and the second positioning post 23 can be easily inserted into the second positioning sleeve 24.

[0037] 8, support columns 25 are fixed at equal intervals to the bottom of the bottom connection plate 191. Rubber friction pads 26 are fixed to the bottom of the support columns 25. This allows the bottom connection plate 191 to be stably positioned.

[0038] Furthermore, the sealing film 3 includes an elastic film or a non-elastic film. In the present invention, the sealing film 3 is preferably a non-elastic film rather than an elastic film, because the non-elastic film can be more easily laminated to the top of the circular kit 2 to form a better sealing environment, while the cost of the non-elastic film is lower.

[0039] The present invention further provides a manufacturing process for the nucleic acid detection kit, the manufacturing process comprising the steps of:

[0040] 1) The injection port 8 is closed with a rubber plug 9, and the nucleic acid extraction reagent and magnetic beads in the circular kit 2 are sealed with a U-shaped aluminum foil film 18. The nucleic acid extraction reagent and magnetic beads are placed in the corresponding storage ports in the circular kit 2, and the magnetic beads may be replaced with a magnetic bead solution. Here, the extraction reagents are a dissolution liquid, a cleaning liquid 1, a cleaning liquid 2, a cleaning liquid 3, and a cleaning liquid 4. The dissolution liquid is contained in a dissolution liquid storage tube e, and the cleaning liquids 1 to 4 are contained in the cleaning liquid storage port 1a, the cleaning liquid storage port 2b, the cleaning liquid storage port 3c, and the cleaning liquid storage port 4d, respectively. 2) A liquid or solid reagent is added into the PCR tube 15, and then the embedded tube assembly 16 is pressed in, and the order of steps 1 and 2 can be reversed. 3) The sealing film 3 is adhered and crimped to the top of the circular kit 2 and the bottom of the top cover 7. If necessary, a desiccant or the like can be placed on the bottom of the top cover 7 in advance. 4) An external gas source is connected, and gas is introduced through the detection hole 27 of the upper cover 7 to detect the hermeticity of the sealing film 3. 5) If no gas leakage is detected, the detection hole 27 in the top cover 7 is blocked with the detection plug 28.

[0041] In a preferred embodiment of the present invention, a desiccant is fixed in the space within the sealing film 3. The desiccant is fixed to the upper cover 7 or the base, and may be located in a position that does not affect the movement of the metal rod 10 and the inclusion of the reagent.

[0042] The PCR tube 15 has an outer wall thickness of 0.1-1.5 mm and a light transmittance of 60% or more so that optical detection is possible.

[0043] The top surface of the U-shaped aluminum foil film 18 does not react with the reagent contained therein, and a PP plastic film is attached to the bottom surface, so that the liquid and solid in the shallow tube 17 can be stored while being in contact with the liquid and solid. The aluminum foil film A163 at the top of the embedded tube body 161 can be in contact with the reagent on one side, and the aluminum foil film B162 at the bottom can be in contact with the reagent on both sides. The order of sealing the embedded tube assembly 16 body is to seal the aluminum foil film B162 at the bottom first, put in the embedded reagent, and then seal the aluminum foil film A163 at the top. The liquid or solid reagent at the bottom is put into the PCR tube 15, and then the bottom surface of the aluminum foil film B162 is washed and the embedded tube assembly 16 is put in. The aluminum foil film B162 at the bottom of the embedded tube assembly 16 and the step at the bottom of the embedded tube assembly 16 are pressed against the top of the PCR tube 15 to seal. Alternatively, the aluminum foil film B162 at the bottom of the embedded tube assembly 16 and the step at the bottom of the embedded tube assembly 16 are fastened to the internal structure of the circular kit 2 at the top of the PCR tube 15 to seal the PCR tube 15 below. The embedded tube assembly 16 achieves sealing of the PCR tube 15 by pressure contact, i.e., structural fastening, welding or bonding. The step at the bottom of the embedded tube assembly 16 may be an arc-shaped step.

[0044] Working principle: When in use, reagents, magnetic beads or magnetic bead solutions are placed into each storage port on the circular kit 2, then the embedded tube assembly 16 is sealed in the PCR tube 15, and the U-shaped aluminum foil membrane 18 is sealed onto the shallow tube 17. After the sealing operation is completed, the sealing membrane 3 is glued onto the circular kit 2, and the lower end of the sealing membrane 3 is pressed onto the circular kit 2 with the latex plastic pressing cap 4. During the pressing process, the lower latex plastic pressing cover 4 is lifted and rotated so that the first positioning sleeve 22 on the lower latex plastic pressing cover 4 is aligned with the first positioning pillar 21 on the lower ring fixed cover 1, and the lower latex plastic pressing cover 4 is moved downward so that the first positioning sleeve 22 is fitted onto the outside of the first positioning pillar 21, completing the positioning work of the lower latex plastic pressing cover 4, and finally inserting the upper end of the first bolt 5 into the first screw hole, rotating the first rotating wheel 198 to rotate the worm 197, thereby causing the worm 197 to rotate the worm wheel 195. The worm wheel 195 rotates the inner toothed ring 192, which in turn rotates the six first gears 194, which in turn rotates the first connecting shaft 193 and rotates the six first bolts 5 simultaneously, causing the first bolts 5 to screw into the first screw holes. As a result, the first bolts 5 pull the lower latex plastic pressing lid 4 downward, and ultimately the lower latex plastic pressing lid 4 presses the sealing film 3 against the circular kit 2.

[0045] Further, the upper cover 7 is attached and the upper end of the sealing film 3 is pressed. The upper cover 7 is lifted, the metal rod 10 on the upper cover 7 is inserted into the metal rod receiving hole 6 in the middle of the circular kit 2, the upper latex plastic pressing cover 12 on the outer side of the sealing film 3 is lifted, and the upper latex plastic pressing cover 12 is attached. The six second positioning sleeves 24 on the upper latex plastic pressing cover 12 are aligned with the six second positioning posts 23 on the circular fixed cover 11, the upper latex plastic pressing cover 12 is moved upward so that the second positioning sleeves 24 are fitted on the outer side of the second positioning posts 23, and finally the second screw hole is fitted on the outer side of the lower end of the second bolt 13. Further, the upper latex plastic pressing cover 12 and the upper circular fixed cover 11 are crimped. Next, the second rotating wheel 205 is rotated to rotate the second connecting shaft 201 fixed to the second rotating wheel 205, thereby rotating one second bolt 13 fixed to the second connecting shaft 201. When the second connecting shaft 201 rotates, the second gear 202 is rotated, and the gear ring 204 is rotated, and the six second gears 202 are rotated simultaneously to rotate the six second bolts 13 simultaneously, whereby the second bolts 13 are screwed into the second screw holes. Furthermore, the upper latex plastic pressing lid 12 is pulled upward until it presses the sealing film 3 against the upper lid 7, thereby completing the sealing operation. After the sealing is completed, the detection plug 28 is opened, and gas is introduced from the detection hole 27 to detect whether there is a gas leak in the sealing film 3. After the detection is completed, the detection plug 28 is closed.

[0046] The present invention can greatly improve the sealing efficiency, is convenient to operate, and can detect gas leakage after sealing is completed, and compared with the conventional detection kit, the sealing quality is greatly improved and the sealing efficiency is improved. In addition, the nucleic acid detection kit provided by the present invention includes a complete process of nucleic acid extraction, nucleic acid amplification and nucleic acid detection, which can ensure the sealed environment of the nucleic acid detection reagent device, reduce the risk of sample cross-contamination, and effectively improve the detection efficiency of nucleic acid detection.

[0047] The above description is merely an embodiment of the present invention, and does not limit the scope of protection of the present invention. Any conversion of equivalent structures or equivalent processes made by those skilled in the art based on the contents of this specification and drawings, or direct or indirect use of other related technical fields, are all within the scope of protection of the present invention.

Claims

1. A nucleic acid detection kit, comprising: The nucleic acid detection kit includes a lower circular fixed cover (1), a circular kit (2) is inserted inside the lower circular fixed cover (1), a sealing film (3) is provided on the top of the circular kit (2), a lower latex plastic pressing cover (4) is provided on the lower end of the sealing film (3), the bottom of the lower latex plastic pressing cover (4) is in pressing contact with the lower end of the sealing film (3), and the bottom of the sealing film (3) is in pressing contact with the top of the circular kit (2), a first bolt (5) is rotatably connected at an equal distance by a bearing to the outside of the lower circular fixed cover (1), and the first bolt (5) is rotatably connected to the outer side of the lower circular fixed cover (1) by a bearing. The circular kit (5) is screwed to the lower latex plastic pressing lid (4) through a first screw hole, a metal rod receiving hole (6) is provided in the center of the circular kit (2), a top lid (7) is provided above the sealing membrane (3), an injection port (8) is provided on one side of the circular kit (2), a rubber plug (9) is tightly fitted into the center of the injection port (8), a metal rod (10) is fixed to the center of the top lid (7), and the lower end of the metal rod (10) is inserted into the metal rod receiving hole (6), an upper circular fixing lid (11) is fixed to the outer wall of the top lid (7), and the sealing membrane (3) is connected to the injection port (8) through a first screw hole (6). An upper latex plastic pressing lid (12) is provided at the upper end of the circular kit (2), the top of the upper latex plastic pressing lid (12) is in pressing contact with the top end of the sealing film (3), and the top of the sealing film (3) is in pressing contact with the bottom of the upper lid (7). A second bolt (13) is rotatably connected to the outer ring of the upper circular fixed lid (11) at an equal distance by a bearing, and the second bolt (13) is screwed into the upper latex plastic pressing lid (12) by a second screw hole. An embedded pipe mounting hole (14) is opened on one side of the circular kit (2), and an embedded pipe mounting hole (14) is opened on the bottom of the circular kit (2). a PCR tube (15) is fixed to the outside of the buried tube mounting hole (14), an buried tube assembly (16) is provided inside the buried tube mounting hole (14), shallow tubes (17) are arranged at equal intervals on the side of the circular kit (2) farther from the PCR tube (15), a U-shaped aluminum foil film (18) is provided on the side of the top of the circular kit (2) closer to the shallow tubes (17), a detection hole (27) is opened on one side of the top cover (7), and a detection plug (28) is tightly fitted and connected to the inner wall of the detection hole (27).

2. The nucleic acid detection kit of claim 1, characterized in that a first connection mechanism (19) is fixed to the bottom of the lower circular fixed lid (1), and the first connection mechanism (19) is used to drive the simultaneous rotation of a first bolt (5), and a second connection mechanism (20) is provided on the top of the upper circular fixed lid (11), and the second connection mechanism (20) is used to drive the simultaneous rotation of a second bolt (13).

3. 3. The nucleic acid detection kit according to claim 2, wherein the embedded tube assembly (16) comprises an embedded tube body (161), an aluminum foil film B (162), and an aluminum foil film A (163), the embedded tube body (161) is provided on the inner wall of the embedded tube mounting hole (14), the aluminum foil film B (162) is heat sealed to the bottom of the embedded tube body (161), the aluminum foil film A (163) is heat sealed to the top of the embedded tube body (161), and the bottom of the aluminum foil film B (162) is pressed into contact with the top of the PCR tube (15).

4. The first connection mechanism (19) includes a bottom connection plate (191), an inner gear ring (192), a first connection shaft (193), a first gear (194), a worm wheel (195), a support block (196), a worm (197), and a first rotating wheel (198). The bottom connection plate (191) is fixed to the bottom of the lower ring fixing cover (1), the inner gear ring (192) is rotatably connected to the lower surface of the bottom connection plate (191) by a bearing, the first connection shaft (193) is fixed to the center of the lower surface of the first bolt (5), and the first gear (194) is fixed to the center of the first connection shaft (193). The nucleic acid detection kit of claim 3, characterized in that each of the first gears (194) is meshed with an internal toothed ring (192), a worm wheel (195) is fixed to the bottom of the internal toothed ring (192), support blocks (196) are symmetrically fixed to one side of the underside of the bottom connecting plate (191), a worm (197) is rotatably connected between the two support blocks (196) by bearings, the worm (197) is meshed with the worm wheel (195), and one end of the worm (197) passes through the support block (196) and is fixed to a first rotating wheel (198).

5. The nucleic acid detection kit of claim 3, characterized in that first positioning pillars (21) are fixed at equal intervals to the top of the lower circular fixing lid (1), first positioning sleeves (22) are fixed at equal intervals to the outer wall of the lower latex plastic pressing lid (4), and the first positioning pillars (21) are inserted into the first positioning sleeves (22).

6. The nucleic acid detection kit according to claim 5, characterized in that the second connection mechanism (20) includes a second connecting shaft (201), a second gear (202), a rotating ring (203), a gear ring (204), and a second rotating wheel (205), the second connecting shaft (201) is fixed to the center of the upper surface of the second bolt (13), the second gear (202) is fixed to the upper end of the second connecting shaft (201), a rotating ring (203) is rotatably connected to the center of the upper surface of the upper circular ring fixing cover (11) by a bearing, a gear ring (204) is fixed to the outer wall of the upper end of the rotating ring (203), all six second gears (202) are meshed and connected to the gear ring (204), and a second rotating wheel (205) is fixed to the top of one second connecting shaft (201) on one side of the upper circular ring fixing cover (11).

7. The nucleic acid detection kit of claim 5, characterized in that second positioning posts (23) are fixed at equal intervals to the outer wall of the upper circular fixed cover (11), second positioning sleeves (24) are fixed at equal intervals to the outer wall of the upper latex plastic pressing cover (12), the second positioning posts (23) are inserted into the second positioning sleeves (24), the first rotating wheel (198) and the second rotating wheel (205) are both plum-shaped, and both the top of the first positioning post (21) and the bottom of the second positioning post (23) are chamfered.

8. The nucleic acid detection kit according to claim 3, characterized in that support columns (25) are fixed at equal intervals to the bottom of the bottom connecting plate (191), and rubber friction pads (26) are fixed to the bottom of the support columns (25).

9. The nucleic acid detection kit according to claim 1 , wherein the sealing film (3) comprises an elastic film or a non-elastic film.

10. A process for producing the nucleic acid detection kit according to any one of claims 1 to 8, comprising the following steps 1) to 5): 1) The injection port (8) is closed with a rubber plug (9), and the nucleic acid extraction reagent and magnetic beads in the circular kit (2) are enclosed with a U-shaped aluminum foil membrane (18). The nucleic acid extraction reagent and magnetic beads are placed in corresponding storage ports in the circular kit (2), and the magnetic beads may be replaced with a magnetic bead solution; 2) adding a liquid or solid reagent into the PCR tube (15), and then pressing the embedded tube assembly (16); the order of steps 1) and 2) can be reversed; 3) The sealing film (3) is glued and crimped to the top of the circular kit (2) and the bottom of the top cover (7); 4) Connect to an external gas source and introduce gas through the detection hole (27) in the top cover (7) to detect the tightness of the sealing film (3); 5) A manufacturing process characterized in that, if no gas leakage is detected, the detection hole (27) in the top cover (7) is blocked with a detection plug (28).

11. A desiccant is fixed in the space within the sealing film (3), and the desiccant is fixed to the top cover (7) or the base, and may be located in a position that does not affect the movement of the metal rod (10) or the inclusion of the reagent. The manufacturing process according to claim 10, wherein the PCR tube (15) has an outer wall thickness of 0.1-1.5 mm and a light transmittance of 60% or more to enable optical detection.

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