Nucleic acid extraction and amplification kit

Abstract

The nucleic acid extraction and amplification kit comprises a sample area, a cracking area, a waste liquid area, a reagent bottle area, a tool area and an amplification detection area which are combined into a whole, the sample area is used for placing a to-be-detected sample, and the cracking area is used for cracking the to-be-detected sample and collecting to-be-detected nucleic acid in the to-be-detected sample; the waste liquid area is used for storing waste liquid; the reagent bottle area is used for storing reaction liquid required by nucleic acid extraction and amplification; the tool area is used for storing tools required for pipetting; the amplification detection area is provided with an eight-connection tube and is used for amplifying and detecting nucleic acid to be detected. The kit is simple in structure, high in integration level, capable of completing detection of 8 * n items (n is the number of fluorescent channels) and position arrangement and placement of all areas at the same time, convenient to operate, capable of keeping balance during placement, simple in structure, small in size, light in weight and suitable for mass production and transportation.

Description

Technical Field

[0001] The utility model belongs to the field of biological detection and relates to a nucleic acid extraction and amplification kit. Background Art

[0002] Nucleic acid extraction and PCR amplification detection are important experimental methods in molecular biology research. Early experiments were all done manually, which took a long time and the experimental results were easily affected by individual differences in experimental personnel. With the development of industrial automation, in the field of experimental science, traditional manual experiments are increasingly replaced by automated equipment. The repeatability and consistency of automated equipment reduce the errors of traditional manual experiments and improve the reliability and efficiency of experiments. The nucleic acid extraction and detection all-in-one machine is a highly integrated compact instrument that performs fully automatic, fully closed, and integrated operations for nucleic acid extraction and amplification. It can highly automate the automation of nucleic acid extraction and amplification detection, shorten the nucleic acid extraction and amplification process, reduce the impact of human factors, enhance the safety and effectiveness of nucleic acid sample preparation, and realize the miniaturization and portability of the device to meet the needs of grassroots or on-site rapid detection. It is the main research direction for the development of new nucleic acid extraction and detection all-in-one machines.

[0003] The nucleic acid extraction and detection machine needs to use a nucleic acid extraction and amplification kit to complete the extraction and detection of nucleic acids. The nucleic acid extraction and amplification kit is a disposable consumable used in the nucleic acid extraction and detection machine. However, the existing nucleic acid extraction and amplification kits have poor integration and are difficult to complete the nucleic acid extraction and detection of complex samples. In addition, the throughput is low and usually only one nucleic acid can be amplified at a time. When there are multiple projects, multiple operations are required, which takes a lot of time. Therefore, it is urgent to find a nucleic acid extraction and amplification kit with a simple structure, small size, and the ability to detect multiple projects at the same time. Utility Model Content

[0004] In order to solve the above problems, the utility model provides a nucleic acid extraction and amplification kit, including a sample area, a lysis area, a waste liquid area, a reagent bottle area, a tool area and an amplification detection area that are integrated into one body. The sample area is used to place the sample to be tested, the lysis area is used for the lysis treatment of the sample to be tested, and the nucleic acid to be tested in the sample to be tested is collected; the waste liquid area is used to store waste liquid; the reagent bottle area is used to store the reaction solution required for nucleic acid extraction and amplification; the tool area is used to store the tools required for pipetting; the amplification detection area is provided with eight connecting tubes for the amplification and detection of the nucleic acid to be tested. The kit has a simple structure and high integration, and can complete the detection of 8*n items (n is the number of fluorescent channels) at the same time. The position setting and placement of each area can take into account the convenience of operation and keep balance when placed. It has a simple structure, small size and light weight, and is suitable for mass production and transportation.

[0005] The nucleic acid extraction and amplification kit provided by the utility model comprises a sample area, a lysis area, a waste liquid area, a reagent bottle area, a tool area and an amplification detection area, wherein the sample area, the lysis area, the waste liquid area, the reagent bottle area, the tool area and the amplification detection area are integrated into an integrated structure; the sample area is used for placing samples to be tested; the lysis area is used for lysis treatment of samples to be tested and collecting nucleic acids to be tested in the samples to be tested; the waste liquid area is used for storing waste liquid; the reagent bottle area is used for storing reaction liquid required for nucleic acid extraction and amplification; the tool area is used for storing tools required for pipetting; and the amplification detection area is used for amplification and detection of nucleic acids to be tested.

[0006] Furthermore, the upper surface of the reagent box is arranged horizontally in a rectangular shape, and the lower part of the reagent box is arranged according to the actual space required by each area.

[0007] In some embodiments, the bottom of the reagent box is arranged at different heights according to the actual space required by each area, wherein the waste liquid area and the tool area are of the same length and longer than other areas, so that the reagent box can be placed stably.

[0008] Furthermore, the amplification detection area is a row of eight connected tubes, and the eight connected tubes are arranged on one side of the reagent kit, and are arranged in sequence along the long sides close to the rectangular upper surface of the reagent kit. The eight connected tubes are pre-installed with nucleic acid detection reagents.

[0009] The eight-tube connection contains amplification reagents for eight different target nucleic acids to be tested, thereby achieving the simultaneous detection of 8*n items (n is the number of fluorescent channels), which means that at least eight items can be tested simultaneously. For example, when dual fluorescent channels are used, 16 items can be tested simultaneously.

[0010] Furthermore, the lysis zone provides space for lysis of the sample to be tested and extraction of the nucleic acid to be tested, and the lysis zone is arranged close to the wide side of the rectangular upper surface of the reagent kit; the waste liquid zone is arranged parallel to the lysis zone; the lysis zone and the waste liquid zone are both trough bodies open upward, and the bottom of the trough body of the lysis zone protrudes downward to form a small space.

[0011] Furthermore, the openings of the cracking zone and the waste liquid zone are covered with a silicone pad and a cover plate, the silicone pad is located below the cover plate, a through hole is provided on the cover plate, and a cross puncture hole is provided on the silicone pad, and the cross puncture hole is located below the through hole.

[0012] The cross-puncture holes on the silicone pad allow the tip to be easily inserted during use and have a certain sealing effect after use. Therefore, the tip can be inserted and withdrawn from the lysis area and waste liquid area multiple times, so there will be no impact even if multiple projects are tested.

[0013] Furthermore, the reagent bottle area is used to store reagent bottles, and the reagent bottles are sealed with aluminum foil.

[0014] Since various reaction reagents are stored in the reagent bottles in advance, they must be strictly sealed, and silicone pads cannot be used to ensure a better sealing effect.

[0015] Furthermore, the reagent bottle area includes a first reagent bottle area and a second reagent bottle area; the first reagent bottle area is arranged in the middle position of the test kit, close to the waste liquid area, and the first reagent bottle area is equipped with a lysis solution reagent bottle, a magnetic bead reagent bottle, a washing solution reagent bottle, a paraffin oil reagent bottle, an amplification reaction initiator reagent bottle and a backup area; the second reagent bottle area is arranged near another wide side of the rectangular upper surface of the test kit, and the second reagent bottle area is equipped with an eluent reagent bottle and a paraffin reagent bottle.

[0016] Nucleic acid extraction and amplification kits are usually disposable consumables that need to be produced and transported in large quantities. They are required to be small in size and light in weight. At the same time, they must be able to maintain balance when placed alone and not easily tip over, so as to prevent the reagent bottles or tools in the kit from falling off or being misplaced. However, the kit contains a variety of reagent bottles with different content specifications, each of which contains reagents of different contents, as well as tips, puncture tips, eight-way tubes, etc. These reagent bottles and tips must not fall off or be misplaced when placed, and must be easy to get when used, and must not be stuck. At the same time, the overall balance of the kit must be maintained and it must not be easily tipped over.

[0017] Therefore, the location of each area in the test kit needs to be considered comprehensively and must be determined through rigorous testing and screening. The placement of each reagent bottle in the reagent bottle area is selected after a large number of repeated operation experiments. It can simplify the pipetting operation steps and avoid errors, while keeping the overall balance of the test kit and preventing it from tipping over. At the same time, the reagent bottle area must be divided into two reagent bottle areas and placed in different positions of the test kit. This can better keep the test kit balanced, while also taking into account the convenience of operation and preventing errors.

[0018] For example, the eluent reagent bottle and the paraffin reagent bottle need to be placed in the second reagent bottle area, separated from the first reagent bottle area. The main reason is that both the eluent and the paraffin need to be heated before use. The eluent needs to be preheated to the same temperature as the pyrolysis area, and the paraffin needs to be heated and melted before use. Therefore, the eluent reagent bottle and the paraffin reagent bottle are placed in the same area and heated together.

[0019] Furthermore, the tool area is located between the first reagent bottle area and the second reagent bottle area, and includes a tip head area and a puncture head area; the tip head area is used to store the tip head, and the tip head is equipped with a filter element; the tip head area is also provided with a backup area for backing up other tip heads or other tools; the puncture head area is used to store the puncture head for piercing the sealing film of the reagent bottle.

[0020] The tool area is located between the first reagent bottle area and the second reagent bottle area, which is beneficial to maintaining the overall balance of the reagent kit and also helps to simplify the pipetting operation steps and prevent errors.

[0021] The tip head described in the utility model is a disposable liquid transfer gun head, which is made of plastic.

[0022] Furthermore, the sample area is located on one side of the eight-tube connection and is a hollow cylindrical area that can be used to place test tubes containing samples to be tested.

[0023] Furthermore, a bracket is provided below the reagent box, and buckles are provided on both sides of the bracket for fixing the reagent box.

[0024] In some embodiments, the support is a semi-enclosed fence surrounding the first reagent bottle area and the tool area, the upper part of the support is connected to the upper surface of the reagent box, the lower part is suspended, the buckles are located on both sides of the support, and one buckle is set on one side. The reagent box is fixed in the nucleic acid extraction and detection integrated machine by two buckles.

[0025] In some embodiments, the nucleic acid extraction and amplification kit provided by the present invention can be made into a low-cost disposable consumable, or can be made into a device that can be sterilized and reused in whole or in part.

[0026] The nucleic acid extraction and amplification kit provided by the utility model has the following beneficial effects:

[0027] 1. It can simultaneously perform amplification detection of 8*n (n is the number of fluorescent channels) types of target nucleic acids;

[0028] 2. The location and placement of each area in the test kit should not only take into account the convenience of operation and prevent errors, but also keep the test kit balanced when placed alone and not prone to tipping over, so as to prevent the reagent bottles or tools in the test kit from falling off and being misplaced;

[0029] 3. Simple structure, small size and light weight, suitable for mass production and transportation. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 A schematic diagram of the structure of the nucleic acid extraction and amplification kit provided in Example 1;

[0031] Figure 2 An exploded view of the nucleic acid extraction and amplification kit provided in Example 1;

[0032] Figure 3 An exploded view of the nucleic acid extraction and amplification kit provided in Example 1 (viewed from a lower angle);

[0033] Figure 4 A top view of the nucleic acid extraction and amplification kit provided in Example 1;

[0034] Figure 5 A front view of the nucleic acid extraction and amplification kit provided in Example 1;

[0035] Figure 6 A rear view of the nucleic acid extraction and amplification kit provided in Example 1;

[0036] Figure 7 A left view of the nucleic acid extraction and amplification kit provided in Example 1;

[0037] Figure 8 A right view of the nucleic acid extraction and amplification kit provided in Example 1;

[0038] Fig. 9 A bottom view of the nucleic acid extraction and amplification kit provided in Example 1;

[0039] Fig.10 This is a schematic diagram of the structure of the puncture head provided in Example 1. DETAILED DESCRIPTION

[0040] The preferred embodiments of the present invention are further described in detail below in conjunction with the accompanying drawings. It should be pointed out that the following embodiments are intended to facilitate the understanding of the present invention and do not have any limiting effect on it. All features disclosed in the embodiments of the present invention, or all steps in the disclosed methods or processes, except for mutually exclusive features and / or steps, can be combined in any way.

[0041] Example 1: Nucleic acid extraction and amplification kit provided by the utility model

[0042] The nucleic acid extraction and amplification kit provided in this embodiment is as follows Figures 1 to 10 As shown, Figure 1 This is a schematic diagram of the overall structure of the nucleic acid extraction and amplification kit. Figure 2 This is an exploded view of the nucleic acid extraction and amplification kit; Figure 3 This is an exploded view of the nucleic acid extraction and amplification kit (viewed from a lower angle); Figure 4 A top view of a nucleic acid extraction and amplification kit; Figure 5 It is a front view of the nucleic acid extraction and amplification kit; Figure 6 This is the rear view of the nucleic acid extraction and amplification kit; Figure 7 This is the left view of the nucleic acid extraction and amplification kit; Figure 8 This is the right view of the nucleic acid extraction and amplification kit; Fig. 9 This is a bottom view of the nucleic acid extraction and amplification kit; Fig.10 It is a schematic diagram of the structure of the puncture head.

[0043] When used, the nucleic acid extraction and amplification kit 1 (hereinafter referred to as kit 1) provided in this embodiment needs to be installed in a nucleic acid extraction and detection all-in-one machine, and cooperate with the nucleic acid extraction and detection all-in-one machine to complete the extraction, amplification and detection of nucleic acids.

[0044] like Figures 1 to 3 As shown, the reagent kit 1 includes a sample area 2, a lysis area 3, a waste liquid area 4, a reagent bottle area 5, a tool area 6 and an amplification detection area 7, which are integrated into an integrated structure; the sample area 2 is used to place the sample to be tested; the lysis area 3 is used for lysis treatment of the sample to be tested and collection of the nucleic acid to be tested in the sample to be tested; the waste liquid area 4 is used to store waste liquid; the reagent bottle area 5 is used to store the reaction solution required for nucleic acid extraction and amplification; the tool area 6 is used to store the tools required for pipetting; and the amplification detection area 7 is used for amplification and detection of the nucleic acid to be tested.

[0045] like Figure 4 and 9 It can be seen that the upper surface 8 of the reagent box 1 is arranged horizontally in a rectangular shape. Figure 4 and 5 The lower part 9 of the reagent box 1 is arranged according to the actual space required by each area, showing different heights. The length of the waste liquid area 4 and the tool area 6 is the same, and is longer than other areas, so that the reagent box 1 can be placed vertically stably.

[0046] like Figure 2 and 3 The amplification detection area 7 is a row of eight connected tubes 10, which are arranged on one side of the reagent kit 1, and are arranged in sequence along a long side 11 of a rectangular upper surface 8 of the reagent kit 1. The eight connected tubes 10 are pre-installed with nucleic acid detection reagents.

[0047] The eight-tube 10 is respectively filled with 8*n (n is the number of fluorescent channels) amplification reagents (lyophilized powder) of different target nucleic acids to be tested, so that 8*n (n is the number of fluorescent channels) items can be detected simultaneously. For example, 16 different amplification primers of target nucleic acids to be tested are respectively filled, two in each tube, each amplification primer is modified with a fluorescent group, and dual fluorescent channels are used for detection.

[0048] like Figures 1 to 9The lysis area 3 is a trough body with an opening facing upward, which provides space for the lysis of the sample to be tested and the extraction of the nucleic acid to be tested. The bottom of the trough body protrudes downward to form a small space 12. When the reagent kit 1 is placed in the nucleic acid extraction and detection integrated machine, the lysis area 3 is located near the upper end of the magnet in the nucleic acid extraction and detection integrated machine. After completing the lysis of the sample and the extraction of the nucleic acid to be tested, slide the magnet to the bottom of the small space 12. The magnetic adsorption effect of the magnet causes the magnetic beads to gather in the small space 12, so that the solution in the lysis area 4 can be more conveniently removed. The lysis area 3 is set close to a wide side 13 of the rectangular upper surface 8 of the reagent kit 1.

[0049] like Figures 1 to 9 The waste liquid area 4 is also a trough body with an opening facing upwards, and is longer than the cracking area 3, so the capacity is larger, and the waste liquid area 4 is arranged in parallel with the cracking area 3. The openings of the cracking area 3 and the waste liquid area 4 are covered with a silicone pad 14 and a cover plate 15, the silicone pad 14 is located below the cover plate 15, and the cover plate 15 is provided with a through hole 16, and the silicone pad 14 is provided with a cross puncture hole 17, and the cross puncture hole 17 is located below the through hole 16. The cross puncture hole 17 of the silicone pad can realize that the tip head can be easily inserted during use, and can have a certain sealing effect after use, so that the tip head can be inserted and withdrawn from the cracking area 3 and the waste liquid area 4 multiple times, so that there will be no impact even if multiple items are tested.

[0050] The reagent bottle area 5 is used to store reagent bottles 18, which are sealed with aluminum foil. Since various reaction reagents are stored in the reagent bottles 18 in advance, they must be strictly sealed, and silicone pads cannot be used to ensure a better sealing effect. Figure 4 , the reagent bottle area 5 includes a first reagent bottle area 19 and a second reagent bottle area 20; the first reagent bottle area 19 is arranged in the middle position of the reagent kit 1, close to the waste liquid area 4, and the first reagent bottle area 19 contains a lysate reagent bottle 21, a magnetic bead reagent bottle 22, a washing liquid reagent bottle 23, a paraffin oil reagent bottle 24, an amplification reaction initiator reagent bottle 25 (hereinafter referred to as the initiator reagent bottle 25) and a backup reagent bottle 26, wherein the washing liquid reagent bottle 23 also includes a first washing liquid reagent bottle 231 and a second washing liquid reagent bottle 232; the second reagent bottle area 20 is arranged near the other wide side 27 of the rectangular upper surface 8 of the reagent kit 1 (such as Figure 4 ), the second reagent bottle area 20 contains an eluent reagent bottle 28 and a paraffin reagent bottle 29. Since the reagent kit 1 is usually a disposable consumable, it needs to be mass-produced and transported, and it is required to be small in size and light in weight. At the same time, it can also maintain balance when placed alone and not easily tip over, so as to prevent the reagent bottles 18 or tools in the reagent kit 1 from falling off or misplacing. However, the reagent kit 1 contains a variety of reagent bottles 18 with different content specifications, each of which contains reagents with different contents, and also has a tip head 30, a puncture head 31 ( Fig.10 , used to pierce the sealed aluminum foil of the reagent bottle), eight-connecting tube 10, etc. These reagent bottles 18 and tip heads 30, puncture heads 31, eight-connecting tube 10, etc. must not only ensure that they will not fall off or be misplaced when placed, but also be easy to get when used, and cannot be stuck. At the same time, it is also necessary to keep the overall balance of the reagent kit 1 and not easy to tip over. Therefore, the setting of the positions of each area in the reagent kit 1 needs to be considered comprehensively from all aspects, and must be determined through strict testing and screening. The placement position of each reagent bottle 18 in the reagent bottle area 5 is selected after a large number of repeated operation experiments. It can best simplify the pipetting operation steps without making mistakes, and at the same time, it can also keep the reagent kit 1 balanced as a whole and prevent it from tipping over, thereby selecting the best placement method. At the same time, the reagent bottle area 5 must be divided into two reagent bottle areas and placed in different positions of the reagent kit 1 for placement, so that the reagent kit 1 can be better kept balanced, while taking into account the convenience of operation and preventing mistakes. For example, the eluent reagent bottle 28 and the paraffin reagent bottle 29 need to be placed in the second reagent bottle area 20, separated from the first reagent bottle area 19. The main reason is that the eluent and paraffin need to be heated before use. The eluent needs to be preheated to the same temperature as the pyrolysis zone (the pyrolysis zone temperature is 65°C, so the eluent needs to be preheated to 65°C in advance), and the paraffin needs to be heated and melted before use. Therefore, the eluent reagent bottle 28 and the paraffin reagent bottle 29 are placed in the same area and heated together.

[0051] like Figure 4 The tool area 6 is located between the first reagent bottle area 19 and the second reagent bottle area 20, and includes a tip head area 32 and a puncture head area 33; the tip head area 32 is used to store the tip head 30, and the tip head 30 is equipped with a filter element, and the tip head 30 is respectively arranged at the positions of holes 301 to 310 in the tip head area 32; the tip head area 32 is also provided with a backup area 34 for backing up other tip heads 30 or other tools; the puncture head area 33 is used to store the puncture head 31 that pierces the sealing film of the reagent bottle. The tool area 6 is located between the first reagent bottle area 19 and the second reagent bottle area 20, which is beneficial to the overall balance of the reagent kit 1, and is also beneficial to simplifying the pipetting operation steps and preventing errors. The tip head 30 is a disposable pipette tip (such as Zhejiang Boyu Biotechnology Co., Ltd., T200 transparent pipette tip Zhejiang Boyu Biotechnology Co., Ltd., T200 transparent pipette tip), made of plastic.

[0052] like Figure 6The sample area 2 is located on one side of the eight-tube connection 10 and is a hollow cylindrical area that can be used to place test tubes 38 containing samples to be tested. A bracket 35 is also provided below the reagent box 1, and buckles 36 are provided on both sides of the bracket 35 for fixing the reagent box 1. The bracket 35 is a fence with a semi-enclosed structure, surrounding the first reagent bottle area 19 and the tool area 6. The upper part of the bracket 35 is connected to the upper surface 8 of the reagent box, and the lower part is suspended. The buckles 36 are located on both sides of the bracket 35, and one buckle 36 is provided on one side. The reagent box 1 is fixed in the nucleic acid extraction and detection integrated machine by two buckles 36.

[0053] Example 2: Process of nucleic acid extraction and amplification

[0054] This embodiment uses the nucleic acid extraction and amplification kit 1 provided in Example 1 to perform nucleic acid extraction and amplification, including the following steps:

[0055] 1. Place the reagent kit 1 in the nucleic acid extraction and detection machine and fix it. Click to start the test according to the prompts. The instrument will perform a self-check and the lysis zone 3 will start heating.

[0056] 2. The liquid transfer gun of the nucleic acid extraction and detection machine takes the puncture head 31 at the puncture head area 33 of the tool area 6 to puncture the film of the reagent bottle in the first reagent bottle area 19;

[0057] 3. Move the tip 30 of the pipette at the hole 301 to the hole in the sample area 2, take the sample, move it to the lysis area 3, and release the sample;

[0058] 4. Move to the lysate reagent bottle 21 to take the lysate, move to the lysate zone 3, and release the lysate;

[0059] 5. Move the pipette to the hole 22 of the magnetic bead reagent bottle, take the magnetic beads, move them to the lysis zone 3, release the magnetic beads, move the pipette to the designated position, return the TIP head to the hole 301 position after the step is completed, start magnetic attraction (through the magnetic force of the magnet, the magnetic beads are gathered in the small space 12 of the lysis zone 3), and stop heating in the lysis zone 3;

[0060] 6. Move the pipette to hole 301, take out the TIP head, move it to the lysis area 3, and transfer the supernatant to the waste liquid area 4;

[0061] 7. Move the pipette to the hole position of the first washing liquid reagent bottle 231, suck the first washing liquid and transfer it to the lysis area 3, release the washing liquid; after the step is completed, transfer the supernatant to the waste liquid area 4;

[0062] 8. Move the pipette to the hole position 232 of the second washing liquid reagent bottle, take the second washing liquid, transfer it to the lysis area 3, and release the second washing liquid; after the step is completed, transfer the supernatant to the waste liquid area 4;

[0063] 9. Return the TIP head to the hole 301 and release the magnetic attraction;

[0064] 10. Move the pipette to the puncture head area 33, take the puncture head 31, puncture the sealing film of the eluent reagent bottle 28 and the eight-tube connection 10, and return the puncture head 31 after the step is completed; move to the hole 302, take the TIP head 30 and move it to the hole position of the eluent reagent bottle 28, take the eluent and put it in each tube of the eight-tube connection 10 (including 101 to 108, used to dissolve the lyophilized powder in each eight-tube connection);

[0065] 11. Move the pipette to the 28th hole of the eluent reagent bottle, take the eluent, move it to the lysis zone 3, and release the eluent; after the steps are completed, move the pipette to the 28th hole of the eluent reagent bottle to clean the tip;

[0066] 12. Move the pipette to hole 302 and return the tip to start magnetic suction;

[0067] 13. Move the pipette to the hole 303 and take the tip to the lysis zone 3 to take the purified nucleic acid, move it to the starter reagent bottle 25 and release it. After the step is completed, move the pipette to the hole 303 and return the tip.

[0068] 14. Move the pipette to the hole 304 to take the tip, move to the hole 25 of the starter reagent bottle to take the nucleic acid, and then move to the second tube 102 in the eight-tube connection 10 to release the nucleic acid; after the steps are completed, move the pipette to the hole 304 to return the tip;

[0069] 15. Repeat step 14 for the tips at the positions of holes 305, 306, 307, 308, 309, and 310 to transfer the nucleic acid to the third tube 103, the fourth tube 104, the fifth tube 105, the sixth tube 106, the seventh tube 107, and the eighth tube 108 in the eight-tube connection 10, respectively. Figure 4 ) (each time the nucleic acid to be tested is transferred to the eight-tube concatenation, the tip must be changed to avoid cross contamination);

[0070] 16. Move the pipette to hole 302 and move the tip to hole 24 of the paraffin oil reagent bottle to take the paraffin oil. Return the tip after releasing the paraffin oil.

[0071] 17. Start collecting the fluorescence in the eight-tube 10;

[0072] 18. Move the pipette to the hole 301, take out the tip, move it to the lysis area 3, suck out the residual liquid, move it to the waste liquid area 4, and return the tip after the step is completed;

[0073] 19. After collecting fluorescence for 36 times (fluorescence needs to be collected continuously for 40 times for each well, and paraffin sealing is taken out from the 36th time), the pipette moves to the puncture head area 33 to take out the puncture head 31, and pierces the paraffin reagent bottle at the hole position 29 of the paraffin reagent bottle; after the step is completed, return the tip;

[0074] 20. Move the pipette to hole 302, take out the tip, move it to hole 29 of the paraffin reagent bottle, take out the paraffin, and transfer it into the eight-tube 10; return the tip after the step is completed;

[0075] 21. Stop heating the paraffin area and wait until the fluorescence collection is completed. The experiment is completed.

[0076] The application of the utility model is not limited to this. For example, the scope of application of the utility model can be expanded according to its environmental protection. Any person skilled in the art can make various changes and modifications without departing from the spirit and scope of the utility model. Therefore, the scope of protection of the utility model shall be based on the scope defined by the claims.

Claims

1. A nucleic acid extraction and amplification kit, characterized in that: The sample area, lysis area, waste liquid area, reagent bottle area, tool area and amplification detection area are integrated into one; the sample area is used to place samples to be tested; The lysis area is used for lysis treatment of the sample to be tested and collection of the nucleic acid to be tested in the sample to be tested; The waste liquid area is used to store waste liquid; The reagent bottle area is used to store the reaction solution required for nucleic acid extraction and amplification; The tool area is used to store the tools needed for pipetting; The amplification detection area is used for amplification and detection of the nucleic acid to be tested; the amplification detection area is a row of eight connected tubes, which are arranged on one side of the test kit and arranged in sequence along the long sides of the rectangular upper surface of the test kit. The eight connected tubes are pre-installed with nucleic acid detection reagents.

2. The nucleic acid extraction and amplification kit according to claim 1, wherein The upper surface of the reagent box is arranged horizontally in a rectangular shape, and the lower part of the reagent box is arranged according to the actual space required by each area.

3. The nucleic acid extraction and amplification kit according to claim 2, wherein: The lysis zone provides space for lysis of the sample to be tested and extraction of the nucleic acid to be tested, and the lysis zone is arranged close to the wide side of the rectangular upper surface of the reagent kit; the waste liquid zone is arranged parallel to the lysis zone; the lysis zone and the waste liquid zone are both trough bodies open upward, and the bottom of the trough body of the lysis zone protrudes downward to form a small space.

4. The nucleic acid extraction and amplification kit according to claim 3, wherein: The openings of the cracking zone and the waste liquid zone are covered with a silica gel pad and a cover plate, wherein the silica gel pad is located below the cover plate, a through hole is provided on the cover plate, and a cross puncture hole is provided on the silica gel pad, wherein the cross puncture hole is located below the through hole.

5. The nucleic acid extraction and amplification kit according to claim 4, characterized in that: The reagent bottle area is used to store reagent bottles, and the reagent bottles are sealed with aluminum foil.

6. The nucleic acid extraction and amplification kit according to claim 5, characterized in that: The reagent bottle area includes a first reagent bottle area and a second reagent bottle area; the first reagent bottle area is arranged in the middle position of the test kit, close to the waste liquid area, and the first reagent bottle area is equipped with a lysis solution reagent bottle, a magnetic bead reagent bottle, a washing solution reagent bottle, a paraffin oil reagent bottle, an amplification reaction initiator reagent bottle and a backup area; the second reagent bottle area is arranged near the other wide side of the rectangular upper surface of the test kit, and the second reagent bottle area is equipped with an eluent reagent bottle and a paraffin reagent bottle.

7. The nucleic acid extraction and amplification kit according to claim 6, characterized in that: The tool area is located between the first reagent bottle area and the second reagent bottle area, and includes a tip head area and a puncture head area; the tip head area is used to store tip heads, and a filter element is installed in the tip head; the tip head area is also provided with a backup area for backing up other tip heads or other tools; the puncture head area is used to store the puncture head for piercing the sealing film of the reagent bottle.

8. The nucleic acid extraction and amplification kit according to claim 1, characterized in that: The sample area is located on one side of the eight-tube connection and is a hollow cylindrical area that can be used to place test tubes containing samples to be tested.

9. The nucleic acid extraction and amplification kit according to claim 8, characterized in that: A bracket is also provided below the test kit, and buckles are provided on both sides of the bracket for fixing the test kit.

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