Nucleic acid detection card box, detection equipment and detection method

Abstract

The invention discloses a nucleic acid detection card box, detection equipment and a detection method, and relates to the technical field of nucleic acid detection. A housing; a table; the magnetic attraction assembly is arranged on the workbench and used for attracting the magnetic beads into the pipettor; a rotating assembly; a pipetting assembly; and the lifting assembly is used for driving the pipetting assembly to lift, so that the pipetting assembly is communicated with or disconnected from the pipettor. The card box comprises a card box upper split body; the lower card box split body is rotationally connected with the upper card box split body, a plurality of detection grooves are formed in the lower card box split body, and magnetic beads are arranged in the detection grooves; the invention discloses a pipettor. In the whole process of transferring the solution by the pipettor, the magnetic beads are attracted by the magnetic attraction assembly in the detection equipment, and air flow is increased through the blowing and attraction action of the pipettor, so that residual liquid in the magnetic beads or other impurities influencing PCR (Polymerase Chain Reaction) can be removed as much as possible, the extracted nucleic acid has fewer impurities compared with the conventional extracted nucleic acid, and the amplification effect is better.

Description

Technical Field

[0001] The present application relates to the field of nucleic acid detection technology, and in particular to a nucleic acid detection card box, detection equipment and detection method. Background Art

[0002] Nucleic acid testing is widely used in various fields of biomedicine, especially in the detection of infectious disease pathogens. The results of nucleic acid testing are an important criterion for diagnosis. The traditional nucleic acid testing process is cumbersome, including sample processing and nucleic acid extraction, PCR amplification, result detection and other steps, which need to be completed in different instruments and equipment and different independent laboratories. This not only places high demands on the experimental personnel and the experimental environment, but also increases the risk of cross-contamination of samples during multiple transfers, thereby affecting the accuracy of the test results. Summary of the invention

[0003] In order to improve the accuracy of the test results, the present application provides a nucleic acid detection card box, a detection device and a detection method.

[0004] On the one hand, the present application provides a nucleic acid detection card box, which adopts the following technical solution:

[0005] A nucleic acid detection cartridge, comprising:

[0006] The card box upper split body is provided with a sample adding port, and a sample adding cover is provided at the sample adding port;

[0007] The card box lower body is rotatably connected to the card box upper body, and a plurality of detection slots are integrally formed in the card box lower body, and at least one of the detection slots is provided with a magnetic bead;

[0008] A pipette is arranged in the upper split body of the card box and is used to transfer the solution in the detection tank to other detection tanks.

[0009] By adopting the above technical scheme, a plurality of detection slots are arranged in the lower body of the card box. During the test, the sample loading cover is opened, the sample is loaded into the detection slot through the sample loading port, and the sample loading cover is closed. During the test, the lower body of the card box is driven to rotate by an external mechanism, and the solution in the detection slot is transferred to other detection slots by a pipette to complete nucleic acid extraction and purification and / or PCR amplification detection. The entire process is carried out inside the closed card box, which effectively prevents aerosol contamination. The magnetic beads in the detection slot can be combined with nucleic acid. During the entire process of transferring the solution by the pipette, the magnetic beads are attracted by the magnetic attraction component in the detection device, and at the same time, the air flow is increased by blowing and suction to remove the residual liquid or other impurities affecting PCR in the magnetic beads as much as possible, so that the extracted nucleic acid has fewer impurities than conventional extraction and the amplification effect is better.

[0010] In addition, in the present application, the upper split body of the card box and the lower split body of the card box constitute the entire integrated structure of the card box, which has a simple structure and is easy to use. More importantly, it is more airtight than a non-integrated structure card box.

[0011] Pipette transfer of samples

[0012] Optionally, the pipette comprises:

[0013] A pipette is slidably disposed in the upper body of the cartridge along the central axis of the cartridge, and one end of the pipette is opened away from the lower body of the cartridge;

[0014] A needle, used to be inserted into the detection slot, the needle is connected to one end of the pipette close to the lower body of the cartridge, and the needle is in communication with the pipette;

[0015] A reset member is disposed in the upper body of the cartridge and is used to drive the pipette to move away from the lower body of the cartridge so as to separate the needle from the detection slot.

[0016] Optionally, a sealing tube is fixedly disposed in the axial direction of the card box in the upper split body of the card box, the pipette is slidably disposed in the sealing tube, and the sealing tube is disposed close to the side wall of the upper split body of the card box.

[0017] Optionally, a first filter element is provided in the pipette.

[0018] Optionally, the card box upper body is provided with at least two positioning planes, and the positioning planes extend along the height direction of the card box upper body.

[0019] Optionally, the detection tank includes one or more of a cleaning tank, a lysis tank, an elution tank, a binding tank and a PCR tank.

[0020] In the second aspect, the present application provides a nucleic acid detection device, which adopts the following technical solution:

[0021] A nucleic acid detection device, comprising:

[0022] Card box;

[0023] chassis;

[0024] A workbench is arranged in the housing, and the workbench is provided with an installation cavity for installing the card box;

[0025] A magnetic suction component, disposed on a workbench, for adsorbing magnetic beads into the pipette;

[0026] A rotating assembly, disposed on the workbench and used to drive the card box to rotate;

[0027] a liquid transfer assembly, used to communicate with the liquid transfer device to achieve liquid aspiration or discharge; and

[0028] The lifting component is connected to the pipetting component and is used to drive the pipetting component to rise and fall so as to connect or disconnect the pipetting component with the pipette.

[0029] By adopting the above technical solution, when in use, open the sample loading cover, add samples to the detection tank through the sample loading port, cover the sample loading cover, and place the card box in the installation cavity of the workbench. During the test, the lower split of the card box is driven to rotate by the rotating component, and the solution in the detection tank is transferred to other detection tanks by a pipette to complete nucleic acid extraction and purification and / or PCR amplification detection. The entire process is carried out inside the card box, which effectively prevents contamination.

[0030] The magnetic beads in the detection tank can bind to the nucleic acid. During the whole process of the pipette transferring the solution, the magnetic beads are attracted by the magnetic suction component in the detection equipment. At the same time, the blowing and suction actions of the pipette and the pipetting component increase the air flow to remove the residual liquid or other impurities that affect PCR in the magnetic beads as much as possible. As a result, the extracted nucleic acid has fewer impurities than conventional extraction (relying only on evaporation) and better amplification effect.

[0031] Optionally, the magnetic attraction component includes a first driving member and a magnetic attraction member for absorbing magnetic beads, the magnetic attraction member is connected to the first driving member, and the first driving member is arranged on the workbench to drive the magnetic attraction member to approach or move away from the card box.

[0032] Optionally, a vertical plate is fixedly installed in the casing, and the pipetting assembly includes a mounting portion, a first air pipe, a second air pipe and a power source. The first air pipe is vertically inserted into the mounting portion, and the second air pipe is slidably inserted into the first air pipe. The power source is used to drive the second air pipe to extend and retract to allow the first air pipe to inhale or exhaust air.

[0033] In a third aspect, the present application provides a nucleic acid detection method, which adopts the following technical solution:

[0034] A nucleic acid detection method comprises the following steps:

[0035] S1. Open the sample loading cover on the surface of the card box, add the sample to the lysate in the detection tank, and close the sample loading cover tightly.

[0036] S2. Load the card box into the testing equipment as required, install it in the installation cavity, click Start, run the experiment, and the testing equipment rotates the lower part of the card box to complete the switching of the hole positions;

[0037] S3, aspirate proteinase K and add it to the lysis tank, heat the lysis liquid for a period of time, and mix it by pipetting with a pipette during the heating process;

[0038] S301, aspirate the magnetic beads and add them into the lysis tank, mix them and let them stand for a while to allow the nucleic acid and the magnetic beads to fully combine;

[0039] S302, sucking all the liquid into the pipette, moving the magnetic suction component of the magnetic suction assembly close to the pipette, adsorbing the magnetic beads, and at the same time, discharging the liquid;

[0040] S303, the pipette is moved into the cleaning tank, the cleaning liquid is sucked, the magnetic suction piece is moved away from the pipette, the magnetic beads are released, and the mixture is mixed by blowing and suction; all the liquid is sucked into the pipette of the pipette, the magnetic suction piece is moved close to the pipette, the magnetic beads are adsorbed, and the liquid is discharged at the same time;

[0041] S4. Transfer the nucleic acid elution solution in the pipette in S3 to the position of the lyophilized pellets of PCR enzyme, pipette and mix thoroughly, and then distribute to different PCR slots.

[0042] In summary, the present application includes at least one of the following beneficial technical effects:

[0043] 1. In the present application, the upper split body and the lower split body of the card box constitute the entire integrated structure of the card box, which has a simple structure, is easy to use, and has a low processing cost. Compared with a non-integrated structure card box, it has a stronger airtightness.

[0044] 2. The magnetic beads in the detection tank can bind to the nucleic acid. During the whole process of the pipette transferring the solution, the magnetic beads are attracted by the magnetic suction component in the detection equipment. At the same time, the blowing and suction action of the pipette and the pipetting component increases the air flow to remove the residual liquid or other impurities that affect PCR in the magnetic beads as much as possible. As a result, the extracted nucleic acid has fewer impurities than conventional extraction (relying only on evaporation) and better amplification effect.

[0045] 3. When in use, open the sample loading cover, add sample into the test tank through the sample loading port, cover the sample loading cover, and place the card box in the installation cavity of the workbench. During the test, the lower part of the card box is driven to rotate by the rotating component, and the solution in the test tank is transferred to other test tanks by the pipette to complete nucleic acid extraction and purification and / or PCR amplification detection. The whole process is carried out inside the card box, which effectively prevents contamination. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 It is a schematic diagram of the overall structure of the card box in the embodiment of the present application;

[0047] Figure 2 This is a schematic diagram of the structure of the lower split body of the card box in the embodiment of the present application;

[0048] Figure 3 is a structural schematic diagram of the lower split body of the card box in another perspective of the embodiment of the present application;

[0049] Figure 4 is a top view of the card box of the embodiment of the present application;

[0050] Figure 5 This is an embodiment of the present application Figure 4 Middle AA section view;

[0051] Figure 6 It is a schematic diagram of the structure of the pipette mainly embodied in the embodiment of the present application;

[0052] Figure 7 It is a schematic diagram of the overall structure of the detection device of the embodiment of the present application;

[0053] Figure 8 It is a schematic diagram of the internal structure of the casing of the detection device according to the embodiment of the present application;

[0054] Fig. 9 This is a schematic diagram of the structure of the magnetic suction component and the card box in the embodiment of the present application;

[0055] Fig.10 It is a cross-sectional view of the detection workbench of the embodiment of the present application, which mainly shows the structure of the rotating assembly;

[0056] Fig.11 This is a schematic diagram of the structure of the rotating seat mainly embodied in the embodiment of the present application;

[0057] Fig.12 This is a schematic diagram of the structure of the liquid transfer assembly mainly embodied in the embodiment of the present application;

[0058] Fig.13 is a cross-sectional view of a first trachea and a second trachea in an embodiment of the present application;

[0059] Fig.14 It is a structural schematic diagram of the lifting assembly mainly embodied in the embodiment of the present application.

[0060] Description of the accompanying drawings: 1. Upper body of the cartridge; 11. Positioning plane; 111. Positioning groove; 12. Sealing tube; 121. End plug; 122. Limiting ring; 2. Lower body of the cartridge; 21. Sealing film; 3. Pipette; 31. Pipette; 311. Boss; 312. First filter element; 32. Needle; 33. Reset member; 4. Sample loading cover; 41. Cover body; 411. Receiving groove; 412. First filter element; 33. Needle; 34. Reset member; 35. Sample loading cover; 36. Cover body; 37. Receiving groove; 38. First filter element; 39. First filter element; 40. First filter element; 41. First filter element; 42. First filter element; 43. First filter element; 44. First filter element; 45. First filter Second filter element; 42, cover plate; 421, air pressure balance hole; 5, detection slot; 51, cleaning slot; 52, lysis slot; 53, elution slot; 54, PCR slot; 55, combination slot; 56, small hole slot; 6, housing; 61, workbench; 611, installation cavity; 612, adsorption slot; 613, threaded hole; 614, positioning piece; 62, bottom plate; 63, side plate; 64, cooling fan; 6 5. Vertical plate; 651. Guide groove; 652. Sliding block; 7. Magnetic suction assembly; 71. First driving member; 72. Magnetic suction member; 8. Transfer assembly; 81. Mounting part; 811. Mounting block; 8111. Guide rod; 812. Mounting plate; 82. First air pipe; 821. Step groove; 8211. Sealing ring; 8212. Pressure ring; 8213. Compression spring; 83. Second air pipe; 841. Power motor; 842. First lead screw; 843. First sliding block; 8431. Guide groove; 9. Rotating assembly; 91. Rotating seat; 911. Positioning groove; 912. Fan-shaped groove; 92. Rotating motor; 93. Driving gear; 94. Gear ring; 95. Toothed belt; 10. Lifting assembly; 101. Guide rail; 102. Second sliding block; 103. Second lead screw; 104. Lifting motor. DETAILED DESCRIPTION

[0061] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only 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 ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0062] It should be noted that all directional indications in the embodiments of the present invention (such as up, down, left, right, front, back, etc.) are only used to explain the relative position relationship, movement status, etc. between the components under a certain specific posture (as shown in the accompanying drawings). If the specific posture changes, the directional indication will also change accordingly.

[0063] In the present invention, unless otherwise clearly specified and limited, the terms "connection", "fixation", etc. should be understood in a broad sense. For example, "fixation" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements, unless otherwise clearly defined. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0064] In addition, if there are descriptions involving "first", "second", etc. in the embodiments of the present invention, the descriptions of "first", "second", etc. are only used for descriptive purposes and cannot be understood as indicating or implying their relative importance or implicitly indicating the number of technical features indicated. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In addition, the meaning of "and / or" appearing in the full text includes three parallel schemes. Taking "A and / or B" as an example, it includes scheme A, or scheme B, or a scheme that satisfies both A and B. In addition, the technical solutions between the various embodiments can be combined with each other, but it must be based on the ability of ordinary technicians in the field to implement. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such a combination of technical solutions does not exist and is not within the scope of protection required by the present invention.

[0065] The embodiment of the present application discloses a card box for nucleic acid detection.

[0066] Reference Figure 1 A nucleic acid detection cartridge, comprising an upper cartridge body 1, a lower cartridge body 2 and a pipette 3. The lower cartridge body 2 is rotatably connected to the upper cartridge body 1, and the cross-sections of the lower cartridge body 2 and the upper cartridge body 1 are circular for easy rotation. The lower cartridge body 2 and the upper cartridge body 1 are an integrally formed shell structure, and the upper cartridge body 1 and the lower cartridge body 2 are open at one end close to each other and closed at the other end away from each other, so that the cartridge forms a relatively sealed shell.

[0067] Reference Figure 1 The upper body 1 of the card box is provided with at least two positioning planes 11, and the positioning planes 11 extend along the height direction of the upper body 1 of the card box. During the test, the card box is installed in the installation cavity of the test instrument, and the installation cavity of the instrument has a positioning plate adapted to the positioning planes 11. The positioning planes 11 abut against the positioning plate, thereby achieving the purpose of limiting the position of the upper body 1 of the card box, and in the process of driving the lower body 2 of the card box to rotate, the position of the upper body 1 of the card box is kept stable.

[0068] In order to improve the sealing performance, a sealing ring is provided at the connection between the upper body 1 and the lower body 2 of the card box, and the sealing ring is an O-type sealing ring.

[0069] Further, see Figure 1 A positioning groove 111 is provided on the positioning plane 11, and the positioning groove 111 is arranged along a direction perpendicular to the axis of the cartridge. The positioning groove 111 can be a V-shaped groove, a T-shaped groove, or a groove of other shapes. In this embodiment, the positioning groove 111 is arranged as a V-shaped groove. A positioning portion for inserting into the positioning groove 111 is provided on the instrument, thereby serving the purpose of limiting the vertical freedom of the cartridge.

[0070] Optionally, refer to Figure 2 and Figure 3 , a plurality of detection slots 5 are integrally formed in the lower body 2 of the cartridge, and the detection slots 5 include one or more of a cleaning slot 51, a lysis slot 52, an elution slot 53 and a PCR slot 54. In this embodiment, the detection slot 5 includes a cleaning slot 51, a binding slot 55, a lysis slot 52, an elution slot 53 and a PCR slot 54, specifically, including an elution slot 53, a lysis slot 52, a binding slot 55, three cleaning slots 51, and a plurality of PCR slots 54, the elution slot 53 is provided with a nucleic acid elution liquid, the lysis slot 52 is provided with a lysis liquid, the binding slot 55 is reserved with an incubation buffer, the cleaning slot 51 is provided with a cleaning liquid, and the PCR slot 54 is a position for different target PCR primer probes. The PCR slot 54 is a circular slot, and protrudes from the bottom of the lower body 2 of the cartridge to form a plurality of cylindrical structures extending downward; the cleaning slot 51 and the binding slot 55 are fan-shaped slots, and protrude from the bottom of the lower body 2 of the cartridge. The detection slots 5 all protrude downward from the bottom of the lower body 2 of the card box, forming a protruding structure corresponding to the detection slots 5, which is compatible with the driving mechanism of the detection device, thereby enabling the driving mechanism of the detection device to drive the lower body to rotate.

[0071] Furthermore, at least one of the detection slots 5 is provided with magnetic beads. In the present embodiment, the detection slot 5 further includes four small hole slots 56 , in which proteinase K, magnetic beads, paraffin oil, and PCR enzyme freeze-dried pellets are respectively provided.

[0072] In order to prevent contamination, a sealing film 21 is provided at the notch of the detection slot 5 (see Figure 5 ), in this embodiment, the sealing film 21 is made of aluminum-plastic film.

[0073] Reference Figure 4 and Figure 5The upper body 1 of the card box is provided with a sample loading port, and a sample loading cover 4 is provided at the sample loading port. A plurality of detection slots 5 are integrally formed in the lower body 2 of the card box; when adding samples, the sample loading cover 4 is opened, the sample solution is sucked by the sample loading device and extended into the upper body 1 of the card box through the sample loading port, the sample solution is added into the detection slot 5, and then the sample loading cover 4 is covered. The sample loading cover 4 includes a cover body 41 and a cover plate 42, and the cover body 41 and the cover plate 42 are integrally formed. When the sample loading cover 4 is covered, the cover body 41 is inserted into the sample loading port, and the cover plate 42 abuts against the top wall of the upper body 1 of the card box, and the size of the cover plate 42 is larger than the sample loading port.

[0074] Reference Figure 4 and Figure 5 A receiving groove 411 is provided in the cover body 41, and a pressure balance hole 421 is provided in the cover plate 42. The pressure balance hole 421 is connected to the receiving groove 411. The setting of the pressure balance hole 421 ensures that the inside of the cartridge can be connected to the outside air, ensuring that the test is carried out normally. In order to prevent contamination, a second filter element 412 is provided at the sample addition port. Specifically, the second filter element 412 is embedded in the receiving groove 411 to effectively prevent aerosol from diffusing from the pressure balance hole 421.

[0075] Reference Figure 4 and Figure 5 The pipette 3 is arranged in the upper body 1 of the cartridge, and is used to transfer the solution in the detection tank 5 to other detection tanks 5. The pipette 3 includes: a pipette 31, a needle 32 and a reset member 33.

[0076] Optionally, a sealing tube 12 is fixedly arranged in the axial direction of the upper body 1 of the cartridge, the top of the sealing tube 12 is open and passes through the top wall of the upper body 1 of the cartridge, the top wall of the upper body 1 of the cartridge refers to the end face of the upper body 1 of the cartridge away from the lower body 2 of the cartridge, that is, during the test, the top wall is the topmost wall of the upper body 1 of the cartridge. The pipette 31 is slidably arranged in the upper body 1 of the cartridge along the central axis of the cartridge, the central axis of the cartridge is the vertical axis when the cartridge is in use, and the end of the pipette 31 away from the lower body 2 of the cartridge is opened.

[0077] Reference Figure 5 and Figure 6 The pipette 31 is slidably disposed in the sealing tube 12. A boss 311 is disposed on the pipette 31 for contacting the inner wall of the sealing tube 12. The boss 311 is circular. In order to increase the sealing effect, the side wall of the boss 311 is adapted to and fits the inner wall of the sealing tube 12.

[0078] Reference Figure 5 and Figure 6The bottom of the sealing tube 12 is provided with an end plug 121, which can be fixed to the sealing tube 12 by welding, bonding or threading, and a through hole for the needle 32 to pass through is provided in the end plug 121. The needle 32 is connected to one end of the pipette 31 close to the lower body 2 of the cartridge, and the needle 32 is connected to the pipette 31. The needle 32 is used to pierce the sealing film 21 and is inserted into the detection slot 5, and the needle 32 is in contact with the inner wall of the through hole.

[0079] Optional, see Figure 5 and Figure 6 The reset member 33 is arranged in the upper body 1 of the cartridge to drive the pipette 31 to move away from the lower body 2 of the cartridge, so that the needle 32 is separated from the detection slot 5. In this embodiment, the reset member 33 is set as a reset spring, which is located in the sealing tube 12 and sleeved on the pipette 31. One end of the reset spring abuts against the boss 311, and the other end abuts against the end plug 121. After the needle 32 is inserted into the detection slot 5 and absorbs the solution, the elastic force of the reset spring can drive the pipette 31 and the needle 32 to automatically reset upward, so that the needle 32 is separated from the detection slot 5.

[0080] Reference Figure 6 A limiting ring 122 for abutting against the boss 311 is provided at the tube mouth of the sealing tube 12. The limiting ring 122 is integrally formed with the upper split body 1 of the card box. Under the action of the reset spring, the pipette 31 has a tendency to move upward. When the boss 311 abuts against the limiting ring 122, the position of the pipette 31 is limited to prevent the pipette 31 from escaping from the sealing tube 12.

[0081] Reference Figure 6 The pipette 31 is provided with a first filter element 312, which is in contact with the inner wall of the pipette 31 and is located at the upper end of the pipette 31 to increase the effective transfer volume of the pipette 31. The effective transfer volume is the volume of the cavity between the first filter element 312 and the bottom of the pipette 31. The first filter element 312 can allow gas to pass through and can block aerosols, effectively preventing the aerosol inside the cartridge from diffusing to the outside of the cartridge, and preventing aerosol contamination.

[0082] The embodiments of the present application also disclose a nucleic acid detection device.

[0083] See also Figure 7-Figure 9 A nucleic acid detection device includes a cartridge, a housing 6, a workbench 61, a magnetic suction assembly 7, and a liquid transfer assembly 8. The workbench 61 is disposed inside the housing 6. The workbench 61 is provided with an installation cavity 611 for installing the cartridge. During the test, the cartridge is installed in the installation cavity 611. The housing 6 includes a bottom plate 62, on which two side plates 63 are vertically fixedly disposed. The workbench 61 is fixedly disposed on the two side plates 63.

[0084] Optional, see Fig. 9, a magnetic suction component 7 is provided on the workbench 61, which is used to absorb the magnetic beads in the pipette 3. The magnetic suction component 7 includes a first driving member 71 and a magnetic suction member 72 for absorbing the magnetic beads, and the magnetic suction member 72 is connected to the first driving member 71. The first driving member 71 is provided on the workbench 61 to drive the magnetic suction member 72 to approach or move away from the cartridge. In this embodiment, the magnetic beads are tiny spherical structures made of magnetic material, which can bind to the nucleic acids in the solution; the magnetic suction member 72 is a magnet, which can absorb the magnetic beads when the magnet approaches the magnetic beads. The first driving member 71 is an air cylinder or an oil cylinder, and the piston rod of the first driving member 71 is fixedly connected to the magnetic suction member 72.

[0085] In order to effectively absorb the magnetic beads, the side wall of the workbench 61 close to the magnetic element 72 is provided with an absorption groove 612 that penetrates the installation cavity 611. When the magnetic beads need to be absorbed, the first driving member 71 extends, driving the magnetic element 72 to move along the absorption groove 612 and approach the pipette 31 in the upper body 1 of the cartridge, thereby absorbing the magnetic beads in the pipette 31. The provision of the absorption groove 612 facilitates the magnetic element 72 to approach the pipette 31, improves the absorption effect on the magnetic beads, and prevents some magnetic beads from falling when the pipette 31 blows and sucks air.

[0086] See also Fig.10 and Fig.11 In order to ensure the stable installation of the card box, a threaded hole 613 is opened on the side wall of the workbench 61 near the positioning plane 11 of the card box. The threaded hole 613 penetrates the installation cavity 611, and two groups of threaded holes 613 are provided. Each group of threaded holes 613 corresponds to a positioning plane 11. The threaded hole 613 is threaded with a positioning member 614. The positioning member 614 can be a jumping bean spring, including a positioning column, a spring and a ball. The ball is embedded in one end of the positioning column near the positioning plane 11. The spring is set in the positioning column to squeeze the ball outward. The ball cooperates with the positioning groove 911 on the positioning plane 11 to achieve the purpose of vertical constraint engagement.

[0087] Among them, see Fig.10 and Fig.11 The detection device further includes a rotating assembly 9, which is disposed on the workbench 61 and is used to drive the cartridge to rotate; specifically, the rotating assembly 9 includes a rotating seat 91, a rotating motor 92, a driving gear 93, a gear ring 94, and a toothed belt 95. The rotating seat 91 is rotatably disposed at the bottom of the workbench 61, and the top wall of the rotating seat 91 is provided with a plurality of positioning grooves 911 for inserting the protruding structure of the PCR slot 54, and the top wall of the rotating seat 91 is provided with two fan-shaped grooves 912 to match the protruding structures corresponding to the cleaning slot 51 and the small hole slot 56, and then after the cartridge lower body 2 is embedded in the rotating seat 91, the rotation of the rotating seat 91 can drive the cartridge lower body 2 to rotate.

[0088] The gear ring 94 is fixedly mounted on the rotating seat 91, the rotating motor 92 is fixedly mounted on the workbench 61, the rotating motor 92 can be a servo motor, the driving gear 93 is fixedly mounted on the output shaft of the rotating motor 92, and the toothed belt 95 is mounted on the driving gear 93 and the gear ring 94, and is respectively meshed with the rotating gear and the gear ring 94.

[0089] Optionally, the detection device is further provided with a temperature control component for heating and controlling the temperature of the cartridge in the installation cavity 611, so as to realize temperature control of the cleavage position, elution position and 12 PCR reaction positions of the cartridge. The temperature control component includes a circuit board and a cooling fan 64 arranged on the side plate 63, and the cooling fan 64 is arranged on the side of the housing 6 away from the workbench 61 for heat dissipation.

[0090] Among them, see Figure 8 and Fig.12 A vertical plate 65 is fixedly arranged in the housing 6, and a liquid transfer assembly 8 is arranged on the vertical plate 65, and is used to communicate with the liquid transfer device 3 to achieve liquid suction or discharge. The liquid transfer assembly 8 includes a mounting portion 81, a first air pipe 82, a second air pipe 83 and a power source. The first air pipe 82 is vertically penetrated on the mounting portion 81, and the second air pipe 83 is slidably penetrated in the first air pipe 82, and one end of the second air pipe 83 extending into the first air pipe 82 is closed.

[0091] Optional, see Fig.12 and Fig.13 The mounting portion 81 includes a mounting block 811 and a mounting plate 812, and the mounting plate 812 is fixedly arranged on the side wall of the mounting block 811. A guide groove 651 is provided on the vertical plate 65, and two guide grooves 651 are arranged in parallel. A slider 652 is slidably penetrated in the guide groove 651, and the mounting block 811 is fixedly arranged on the slider 652.

[0092] The power source is used to drive the second air pipe 83 to extend and retract so that the first air pipe 82 can inhale or exhaust air. In this embodiment, the power source is a power motor 841 and a first screw rod 842. The power motor 841 is fixedly arranged on the mounting plate 812, and the first screw rod 842 is coaxially fixedly connected to the output shaft of the power motor 841. The first screw rod 842 is inserted into the second air pipe 83, and the length of the second air pipe 83 is greater than the length of the first screw rod 842. A first sliding block 843 is threadedly connected to the first screw rod 842, and the first sliding block 843 is fixedly connected to the top of the second air pipe 83. Both ends of the first sliding block 843 are provided with guide grooves 8431 in the vertical direction, and the guide grooves 8431 are semicircular arc structures. The top wall of the mounting block 811 is vertically fixedly provided with two guide rods 8111, and the guide rods 8111 are adapted to the guide grooves 8431, and the first sliding block 843 can slide along the guide rods 8111.

[0093] When it is necessary to transfer the solution or blow the magnetic beads, the power motor 841 drives the first screw rod 842 to rotate, the first screw rod 842 drives the first sliding block 843 to move along the guide rod 8111, and the first sliding block 843 drives the second air tube 83 to rise and fall, so that positive pressure or negative pressure is formed in the first air tube 82. When the first air tube 82 is connected to the pipette 31 of the pipette 3, the solution is absorbed or released, and the magnetic beads adsorbed on the side wall of the pipette 31 are blown and sucked. The flow of air accelerates the drying of the residual water on the surface of the magnetic beads, thereby removing the residual liquid or other impurities that affect PCR in the magnetic beads as much as possible.

[0094] Alternatively, see Fig.12 and Fig.13 The first air pipe 82 is provided with a step groove 821, and a sealing ring 8211 is provided in the step groove 821. The sealing ring 8211 can be made of rubber. The sealing ring 8211 is sleeved on the second air pipe 83 to seal the gap between the first air pipe 82 and the second air pipe 83. A pressing ring 8212 is provided on the sealing ring 8211. The bottom wall of the mounting block 811 is provided with a pressing groove in the vertical direction. A compression spring 8213 is provided in the pressing groove. One end of the compression spring 8213 abuts against the bottom wall of the pressing groove, and the other end abuts against the compression ring 8212. Under the action of the compression spring 8213, the sealing ring 8211 is pressed tightly against the step groove 821, so as to prevent the sealing ring 8211 from shifting during the expansion and contraction process of the second air pipe 83, thereby improving the sealing effect.

[0095] Alternatively, see Fig.14 A lifting assembly 10 is provided on the vertical plate 65 , and the lifting assembly 10 is connected to the pipetting assembly 8 , and is used to drive the pipetting assembly 8 to rise and fall, so that the pipetting assembly 8 is connected to or disconnected from the pipette 3 .

[0096] Specifically, the lifting assembly 10 includes a guide rail 101, a second sliding block 102, a second screw rod 103 and a lifting motor 104. The lifting motor 104 is fixedly arranged on the vertical plate 65. The second screw rod 103 is coaxially fixedly connected with the output shaft of the lifting motor 104. The second sliding block 102 is threadedly sleeved on the second screw rod 103. The second sliding block 102 is fixedly connected to the slider 652, and the second sliding block 102 is slidably arranged on the guide rail 101. The lifting motor 104 drives the second screw rod 103 to rotate, and the second screw rod 103 drives the second sliding block 102 to move along the guide rail 101, thereby driving the slider 652 and the mounting portion 81 to rise and fall, thereby achieving the purpose of driving the entire pipetting assembly 8 to rise and fall, so as to facilitate the connection or disconnection of the first air pipe 82 and the pipette 31.

[0097] The implementation principle of the embodiment of the present application is as follows: when in use, open the sample loading cover 4, add sample to the detection slot 5 through the sample loading port, cover the sample loading cover 4, and place the card box in the installation cavity 611 of the workbench 61. During the test, the lower body 2 of the card box is driven to rotate by the rotating component 9, and the solution in the detection slot 5 is transferred to other detection slots 5 by the pipette 3 to complete the nucleic acid extraction purification and / or PCR amplification detection. The whole process is carried out inside the card box to effectively prevent contamination. The magnetic beads in the detection slot 5 can be combined with nucleic acids. During the whole process of the pipette 3 transferring the solution, the magnetic beads are attracted by the magnetic suction component 7 in the detection device. At the same time, the blowing and suction action of the pipette 3 and the pipetting component 8 increases the air flow to remove the residual liquid or other impurities that affect PCR in the magnetic beads as much as possible, so that the extracted nucleic acid has fewer impurities than conventional extraction (relying only on evaporation) and the amplification effect is better.

[0098] The embodiments of the present application also disclose a method for nucleic acid detection.

[0099] A nucleic acid detection method comprises the following steps:

[0100] S1, open the sample loading cover 4 on the surface of the cartridge, add the sample to the lysate in the detection tank 5, and cover the sample loading cover 4 tightly;

[0101] Turn on the instrument and align the label on the test kit with the scanner, scan, retrieve the experimental process and enter the sample information.

[0102] S2. Load the card box into the testing device as required, that is, install it in the installation cavity 611, click Start, run the experiment (hereinafter the instrument automatically runs), and the testing device rotates the lower split body 2 of the card box to complete the switching of the hole positions.

[0103] S3, aspirate proteinase K and add it into the lysis tank 52, heat the lysis liquid for a period of time, and mix it by pipetting with the pipette 3 during the heating process.

[0104] S301, aspirate the magnetic beads and add them into the lysis tank 52, mix them and let them stand for a period of time to allow the nucleic acid and the magnetic beads to fully combine;

[0105] S302, suck all the liquid into the pipette 31, move the magnetic attraction member 72 of the magnetic attraction assembly 7 close to the pipette 31, absorb the magnetic beads, and at the same time, discharge the liquid;

[0106] S303, the pipette 3 is moved into the cleaning tank 51, the cleaning liquid is sucked, the magnetic suction member 72 is moved away from the pipette 3, the magnetic beads are released, and the mixture is mixed by blowing and suction; all the liquid is sucked into the pipette 31 of the pipette 3, the magnetic suction member 72 is moved close to the pipette 3, the magnetic beads are adsorbed, and the liquid is discharged at the same time;

[0107] Specifically, S3031, the pipette 3 is moved into the cleaning tank 51 position I, the cleaning liquid I is sucked, the magnetic suction member 72 is moved away from the pipette 3, the magnetic beads are released, and the mixture is mixed by blowing and suction; all the liquid is sucked into the pipette 31 of the pipette 3, the magnetic suction member 72 is moved close to the pipette 3, the magnetic beads are adsorbed, and the liquid is discharged at the same time;

[0108] S3032, the pipette 3 is moved into the cleaning position II of 51, the cleaning liquid II is sucked, the magnetic suction part 72 is moved away from the pipette 3, the magnetic beads are released, and the mixture is mixed by blowing and suction; all the liquid is sucked into the pipette 31, the magnetic suction part 72 is moved close to the pipette 31, the magnetic beads are adsorbed, and the liquid is discharged at the same time;

[0109] S3033, the sampling needle is moved into the cleaning position III 51, the cleaning liquid III is sucked in, the magnetic suction part 72 is moved away from the sampling needle, the magnetic beads are released, and the mixture is mixed by blowing and suction; all the liquid is sucked into the pipette 31, the magnetic suction part 72 is moved close to the pipette 31, the magnetic beads are adsorbed, and the liquid is discharged at the same time;

[0110] Among them, cleaning tank 51 position I, cleaning tank 51 position II and cleaning tank 51 position III are different cleaning tanks 51 positions.

[0111] S304, the pipette 3 continues to blow and suck for a period of time to fully dry the magnetic beads and avoid the residue of ethanol and other impurities to the greatest extent;

[0112] S305, the pipette 3 is moved into the elution tank 53, the nucleic acid elution liquid is sucked in, the magnetic suction part 72 is moved away from the pipette 31, the magnetic beads are released, and the mixture is mixed by blowing and suction; all the liquid is sucked into the pipette 31, the magnetic suction part 72 is moved close to the pipette 31, and the magnetic beads are adsorbed. In the following steps, the magnetic beads are always adsorbed in the tube wall of the pipette 31.

[0113] S4. Move the nucleic acid elution solution in the pipette 31 in the previous step to the position of the PCR enzyme freeze-dried beads, pipette and mix thoroughly, and then dispense into different PCR tanks 54.

[0114] S5. Absorb paraffin oil and add liquid paraffin oil into each PCR tank 54 to achieve a sealing effect during the amplification process.

[0115] S6. Run the PCR program and detect the fluorescence value in real time.

[0116] S7. Give the test results based on the test data analysis and automatically generate a test report.

[0117] S8. After the experiment, take out the card box, put it into a ziplock bag, seal it, and dispose of it according to the prescribed treatment method.

[0118] The whole process uses the magnetic suction component 7 to absorb the magnetic beads, and at the same time increases the air flow through the blowing and suction action of the pipette 3 to remove as much residual liquid or other impurities that affect PCR as possible from the magnetic beads, so that the extracted nucleic acid has fewer impurities than conventional magnetic bead extraction (relying only on evaporation) and has a better amplification effect.

[0119] The above are all preferred embodiments of the present application, and the protection scope of the present application is not limited thereto. Therefore, any equivalent changes made according to the structure, shape, and principle of the present application should be included in the protection scope of the present application.

Claims

1. A nucleic acid detection cartridge, characterized in that: The card box comprises: The card box upper body (1) is provided with a sample loading port, and a sample loading cover (4) is provided at the sample loading port; A card box lower body (2), the card box lower body (2) being rotatably connected to the card box upper body (1), the card box lower body (2) being integrally formed with a plurality of detection slots (5), at least one of the detection slots (5) being provided with a magnetic bead; A pipette (3), the pipette (3) being arranged in the upper body (1) of the cartridge and being used for transferring the solution in the detection tank (5) to other detection tanks (5).

2. A nucleic acid detection cartridge according to claim 1, characterized in that: The pipette (3) comprises: A pipette (31) is slidably disposed in the upper body (1) of the card box along the central axis of the card box, and the pipette (31) is disposed at an end opening away from the lower body (2) of the card box; A needle (32) is used to be inserted into the detection slot (5), the needle (32) is connected to one end of the pipette (31) close to the lower body (2) of the cartridge, and the needle (32) is in communication with the pipette (31); A reset member (33) is arranged in the upper body (1) of the cartridge and is used to drive the pipette (31) to move in a direction away from the lower body (2) of the cartridge so as to allow the needle (32) to be separated from the detection slot (5).

3. A nucleic acid detection cartridge according to claim 2, characterized in that: A sealing tube (12) is fixedly arranged in the axial direction of the card box in the upper body (1) of the card box, and the pipette (31) is slidably arranged in the sealing tube (12). The sealing tube (12) is arranged close to the side wall of the upper body (1) of the card box.

4. A nucleic acid detection cartridge according to claim 2, characterized in that: A first filter element (312) is arranged in the pipette (31).

5. A nucleic acid detection cartridge according to claim 1, characterized in that: The card box upper body (1) is provided with at least two positioning planes (11), and the positioning planes (11) extend along the height direction of the card box upper body (1).

6. A nucleic acid detection cartridge according to claim 1, characterized in that: The detection tank (5) includes one or more of a cleaning tank (51), a lysis tank (52), an elution tank (53), a binding tank (55) and a PCR tank (54).

7. A nucleic acid detection device, characterized in that: include: The card box according to any one of claims 1 to 6; Housing (6); A workbench (61) is arranged inside the housing (6), and the workbench (61) is provided with an installation cavity (611) for installing the card box; A magnetic attraction component (7), arranged on a workbench (61), and used for absorbing magnetic beads in the pipette (3); A rotating assembly (9), arranged on the workbench (61) and used for driving the card box to rotate; a liquid transfer assembly (8), used for communicating with the liquid transfer device (3) to achieve liquid aspiration or discharge; and A lifting component (10) is connected to the pipetting component (8) and is used to drive the pipetting component (8) to rise and fall so as to connect or disconnect the pipetting component (8) with the pipette (3).

8. A nucleic acid detection device according to claim 7, characterized in that: The magnetic attraction component (7) comprises a first driving member (71) and a magnetic attraction member (72) for absorbing magnetic beads, wherein the magnetic attraction member (72) is connected to the first driving member (71), and the first driving member (71) is arranged on the workbench (61) and is used to drive the magnetic attraction member (72) to approach or move away from the card box.

9. A nucleic acid detection device according to claim 8, characterized in that: A vertical plate (65) is fixedly arranged inside the housing (6), and the pipetting assembly (8) comprises a mounting portion (81), a first air pipe (82), a second air pipe (83) and a power source, wherein the first air pipe (82) is vertically penetrated on the mounting portion (81), and the second air pipe (83) is slidably penetrated inside the first air pipe (82), and the power source is used to drive the second air pipe (83) to extend and retract so that the first air pipe (82) can inhale or exhaust air.

10. A detection method based on the nucleic acid detection device according to any one of claims 7 to 9, characterized in that: The following steps are involved: S1, open the sample loading cover (4) on the surface of the cartridge, add the sample to the lysate in the detection tank (5), and tightly cover the sample loading cover (4); S2. Load the card box into the testing device as required, install it in the installation cavity (611), click Start, run the experiment, and the testing device rotates the lower part (2) of the card box to complete the switching of the hole positions; S3, draw proteinase K and add it into the lysis tank (52), heat the lysis liquid for a period of time, and mix it by pipetting with a pipette (3) during the heating process; S301, aspirate the magnetic beads and add them into the lysis tank (52), mix them and let them stand for a period of time to allow the nucleic acid and the magnetic beads to fully combine; S302, sucking all the liquid into the pipette (31), moving the magnetic attraction member (72) of the magnetic attraction assembly (7) close to the pipette (31), adsorbing the magnetic beads, and at the same time, discharging the liquid; S303, the pipette (3) is moved into the cleaning tank (51), the cleaning liquid is sucked, the magnetic suction part (72) is moved away from the pipette (3), the magnetic beads are released, and the mixture is mixed by blowing and suction; all the liquid is sucked into the pipette (31) of the pipette (3), the magnetic suction part (72) is moved close to the pipette (3), the magnetic beads are absorbed, and the liquid is discharged at the same time; S4, transfer the nucleic acid elution solution in the pipette (31) in S3 to the position of the PCR enzyme freeze-dried pellets, pipette and mix thoroughly, and then dispense into different PCR tanks (54).

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