A sealed integrated device for magnetic bead detection and its application.

Abstract

The application relates to a sealed integrated device for magnetic bead method nucleic acid detection and application thereof, the integrated device comprising a card box, a pipettor and a telescopic elastic sealing cover, the card box comprising an upper cover, a sealing gasket and integrated reagent storage tubes, the integrated reagent storage tubes being sequentially fixedly connected on a connecting body by opening parts of several test tubes, detection reagents being previously filled into different test tubes of the integrated reagent storage tubes, and then the sealing gasket and the upper cover being covered to realize reagent sealing in the card box; the lower part of the telescopic elastic sealing cover being detachably and sealingly connected with the upper part of the card box, and the pipettor being arranged in the telescopic elastic sealing cover. The sealed integrated device can be filled with reagents in advance, automatically operated in a mechanical mode, realizes POCT detection, has a wide application range, is convenient to upgrade to micro-fluidic chip detection, has a simple structure, is easy to match with existing detection facilities, and can be rapidly popularized and applied.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological nucleic acid detection, and particularly relates to a sealed integrated device for nucleic acid detection by a magnetic bead method and application thereof. BACKGROUND

[0002] In recent years, point of care testing (POCT) technology has developed rapidly, and the proportion of POCT in medical testing is also increasing. POCT integrates various professional detection technologies into a very small machine, and the operator can be a non-professional tester, and the detection location is not limited. POCT has a very wide application prospect in the rapid and timely detection of biological detection, disease pathogen detection and food safety monitoring.

[0003] For example, in the detection of disease pathogens, the detection methods of pathogens mainly include biochemical detection, immunodetection and molecular detection. The molecular detection directly detects the nucleic acid of the pathogen, has a higher detection window period than immunodetection, and has the characteristics of high sensitivity and high accuracy. Nucleic acid detection technology is more familiar to the public, and plays an important role in the diagnosis of cases. At present, the nucleic acid detection method mainly collects samples by professional personnel, and then transports the samples to a central laboratory for centralized detection. The sample transportation also needs to maintain cold chain transportation, which increases the difficulty and cost of sample delivery, and also brings great pressure to the testing center. The central laboratory requires that sample pretreatment, reagent preparation and detection analysis be distributed in different rooms to prevent contamination. The strict requirements make it difficult for many primary hospitals to establish a PCR laboratory, and the detection efficiency is greatly reduced.

[0004] In order to improve the efficiency of pathogen detection, the best way is to establish a POCT system, which requires a device that can realize the full sealing of the nucleic acid extraction and amplification detection of clinical samples, and break away from the dependence on the PCR central laboratory, and expand the application range of nucleic acid detection. It is necessary to highly concentrate the operation steps and reagent test tubes of the conventional PCR laboratory and other consumables together to form a device, and to operate through an automatic device. However, there are few such complete devices at present. SUMMARY

[0005] In view of the deficiencies of the prior art, the application provides a sealed integrated device for nucleic acid detection by a magnetic bead method, which comprises a cartridge and a matched pipettor. The cartridge is sealedly connected with the pipettor by a telescopic elastic sealing cover, so that the pipettor needle is sealed from the outside world when moving between the test tubes, and neither gas nor liquid can leak to the outside world. The device can automatically and non-pollutingly realize the sample lysis, magnetic attraction, washing, elution and other steps of the nucleic acid extraction by the magnetic bead method by cooperating with the matched instrument, and finally completes the amplification detection of the nucleic acid.

[0006] In order to achieve the above object, the application provides the following technical scheme: a sealed integrated device for magnetic bead method nucleic acid detection, the integrated device comprising a cartridge, a pipette and a telescopic elastic sealing cover, the cartridge comprising a cover, a sealing gasket and an integrated reagent storage tube, the integrated reagent storage tube being composed of a plurality of open ends of test tubes fixedly connected in sequence on a connecting body, the detection reagent being preloaded into different test tubes of the integrated reagent storage tube, and then the sealing gasket and the cover being covered to realize sealing of the reagent in the cartridge; the cover being provided with a circular opening corresponding to the position of each test tube; the pipette being composed of a needle, a pipette tube and a filter layer; the needle being connected to the rear end of the pipette tube, and the filter layer being fixed to the rear end of the pipette tube; the lower part of the telescopic elastic sealing cover being detachably and sealingly connected to the upper part of the cartridge, so that all the circular openings are covered in the telescopic elastic sealing cover; and the pipette being arranged in the telescopic elastic sealing cover, wherein the rear part of the pipette tube is sealingly connected to the upper part of the telescopic elastic sealing cover.

[0007] Preferably, the telescopic elastic sealing cover is composed of an upper interface, a lower interface and a body, the body being made of a gas and water impermeable elastic material, including silica gel or rubber; the pipette being sealingly connected to the telescopic elastic sealing cover through the upper interface; the upper part of the cartridge being connected to the telescopic elastic sealing cover through the lower interface; and the shape of the body including a flat plate type triangular cylinder, a folded triangular cylinder and an organ type triangular cylinder.

[0008] Further, a microfluidic chip is arranged at the end of the connecting body of the integrated reagent storage tube.

[0009] Further, the part of the microfluidic chip for fluorescence collection is arranged outside the area covered by the telescopic elastic sealing cover.

[0010] Further, the plurality of test tubes of the integrated reagent storage tube are arranged in a straight line, a circle or an array.

[0011] Further, the rear end of the pipette tube is connected to an adapter of a matching machine in a threaded, buckled or tightly fitted manner.

[0012] Further, the pipette further comprises a rubber sleeve wrapped outside the needle, the top end of the rubber sleeve being arc-shaped, corresponding to the circular opening and having a diameter greater than that of the circular opening.

[0013] Further, the sealing gasket is flat or concave in shape corresponding to the position of each test tube opening.

[0014] Further, the rubber sleeve and the sealing gasket are made of a material that is elastic and can be pierced by the needle, including rubber and silica gel.

[0015] Further, the test tubes of the integrated reagent storage tube are in turn: a magnetic bead tube, a lysis solution tube, a washing solution one tube, a washing solution two tube one, a washing solution two tube two, an eluent tube, and a PCR amplification reagent tube.

[0016] Further, when the microfluidic chip is not pre-embedded with PCR amplification reagents, the test tubes of the integrated reagent storage tube are in turn: a magnetic bead tube, a lysis solution tube, a washing solution one tube, a washing solution two tube one, a washing solution two tube two, an eluent tube, and a PCR amplification reagent tube.

[0017] Further, when the microfluidic chip is pre-embedded with PCR amplification reagents, the test tubes of the integrated reagent storage tube are in turn: a magnetic bead tube, a lysis solution tube, a washing solution one tube, a washing solution two tube one, a washing solution two tube two, and an eluent tube.

[0018] Further, the card box, the pipette, and the telescopic elastic sealing cover are disposable.

[0019] Further, on the card cover of the card box, a split cover is arranged at the position corresponding to the circular opening of the lysis solution tube, and the split cover penetrates through the sealing gasket layer.

[0020] The application also provides a detection method based on the magnetic bead method nucleic acid extraction and amplification.

[0021] S1, the adapter is connected with the pipette, and then the adapter is moved above the lysis solution tube, the pipette is moved downward, the needle continues to move downward, penetrates the sealing gasket, enters the lysis solution tube, the air pump drives the pipette to suck all the reagents, then the pipette is lifted upward, the needle moves out of the lysis solution tube, and the sealing gasket rebounds to seal the lysis solution tube;

[0022] S2, the pipette is moved above the magnetic bead tube, is moved downward, the needle enters the magnetic bead tube, the air pump drives the pipette to suck all the liquid in the magnetic bead tube; the air pump is controlled to be pumped several times, the magnetic beads are mixed with the liquid; then the pipette is lifted upward, and the needle moves out of the magnetic bead tube;

[0023] S3, the magnetic bead tube is heated as needed, and after stopping heating, magnetic attraction is performed at a position close to the bottom on the side of the magnetic bead tube for 1-5 minutes;

[0024] S4, the pipette is moved downward, the needle enters the magnetic bead tube, the air pump drives the pipette to suck all the reagents, then the pipette is lifted upward, the needle moves out of the magnetic bead tube, and the magnetic force is removed;

[0025] S5, move the pipette to above the lysis solution tube, move downward, the needle enters the lysis solution tube, the air pump pushes the air, drives the pipette to hit the liquid in it into the lysis solution tube; then the pipette is lifted upward, the needle moves out of the lysis solution tube;

[0026] S6, move the pipette to above the first washing liquid tube, move downward, the needle enters the first washing liquid tube, the air pump drives the pipette to suck all the reagents, then the pipette is lifted upward, the needle moves out of the first washing liquid tube;

[0027] S7, move the pipette to above the magnetic bead tube, move downward, the needle enters the magnetic bead tube, the air pump pushes the air, drives the pipette to hit the liquid in it into the magnetic bead tube, control the air pump to push several times, mix the magnetic beads and the liquid, magnetize at the position close to the bottom of the side of the magnetic bead tube for 1-5 minutes, the air pump pumps the air, the waste liquid is sucked into the pipette, then the pipette is lifted upward, the needle moves out of the magnetic bead tube, and the magnetic force is removed;

[0028] S8, move the pipette to above the first washing liquid tube, move downward, the needle enters the first washing liquid tube, the air pump pushes the air, drives the pipette to hit the waste liquid in it into the first washing liquid tube, then the pipette is lifted upward, the needle moves out of the first washing liquid tube;

[0029] S9, move the pipette to above the first washing liquid tube, move downward, the needle enters the first washing liquid tube, the air pump drives the pipette to suck all the reagents, then the pipette is lifted upward, then the pipette is lifted upward, the needle moves out of the first washing liquid tube;

[0030] S10, a) when the second washing liquid contains alcohol, move the pipette to above the magnetic bead tube, move downward, the needle enters the magnetic bead tube, the air pump pushes the air, drives the pipette to hit the liquid in it into the magnetic bead tube, control the air pump to push several times, mix the magnetic beads and the liquid, magnetize at the position close to the bottom of the side of the magnetic bead tube for 1-5 minutes, the air pump pumps the air, the waste liquid is sucked into the pipette, then the pipette is lifted upward, the needle moves out of the magnetic bead tube, and the magnetic force is removed;

[0031] b) when the second washing liquid does not contain alcohol, move the pipette to above the magnetic bead tube, move downward, the needle enters the magnetic bead tube, the air pump pushes the air, drives the pipette to hit the liquid in it into the magnetic bead tube, magnetize at the position close to the bottom of the side of the magnetic bead tube for 1-5 minutes, then the air pump pumps the air, the waste liquid is sucked into the pipette, then the pipette is lifted upward, the needle moves out of the magnetic bead tube, and the magnetic force is kept;

[0032] S11, move the pipette to above the first washing liquid tube, move downward, the needle enters the first washing liquid tube, the air pump pushes the air, drives the pipette to hit the waste liquid in it into the first washing liquid tube, then the pipette is lifted upward, the needle moves out of the first washing liquid tube;

[0033] S12. Move the pipette above the second washing solution tube and move it downwards. The needle enters the second washing solution tube. The air pump draws air to drive the pipette to draw up all the reagents. Then, the pipette is lifted upwards and the needle is removed from the second washing solution tube.

[0034] S13, a) When the washing solution contains alcohol, move the pipette above the magnetic bead tube, move it downwards, insert the needle into the magnetic bead tube, and use the air pump to push air, which will drive the pipette to inject the liquid into the magnetic bead tube. Control the air pump to pump and push several times to mix the magnetic bead with the liquid. Magnetic attraction is performed on the side of the magnetic bead tube near the bottom for 1-5 minutes. The air pump is then used to draw the waste liquid into the pipette. Then the pipette is lifted upwards, the needle is removed from the magnetic bead tube, and the magnetic force is removed.

[0035] b) When the washing solution does not contain alcohol, move the pipette above the magnetic bead tube, move it downwards, the needle enters the magnetic bead tube, the air pump pushes air, driving the pipette to inject the liquid into the magnetic bead tube, then the air pump draws air to suck the waste liquid into the pipette, then the pipette is lifted upwards, the needle is removed from the magnetic bead tube, and the magnetism is removed.

[0036] S14. Move the pipette above the second washing liquid tube and move it downwards. The needle enters the second washing liquid tube. The air pump pushes air, which drives the pipette to flow the waste liquid into the second washing liquid tube. Then the pipette is lifted upwards and the needle is removed from the second washing liquid tube.

[0037] S15, a) When the washing solution contains alcohol, move the pipette above the magnetic bead tube, move it downwards, insert the needle into the magnetic bead tube, heat the magnetic bead tube to 70°C for half a minute, then lift the pipette upwards and remove the needle from the magnetic bead tube.

[0038] b) This step is omitted if the second washing solution does not contain alcohol;

[0039] S16. Move the pipette above the eluent tube, move it downwards, and the needle enters the eluent tube. The air pump draws air to drive the pipette to draw up all the reagents. Then, lift the pipette upwards and remove the needle from the eluent tube.

[0040] S17. Move the pipette above the magnetic bead tube and move it downwards. Insert the needle into the magnetic bead tube. Use the air pump to push air, which will cause the pipette to inject the elution solution into the magnetic bead tube. Control the air pump to pump and push several times to mix the magnetic bead with the liquid. Heat the magnetic bead tube while controlling the air pump to pump and push several times. After heating, hold the magnetic bead tube near the bottom of the side for 1-5 minutes. Use the air pump to draw air into the pipette containing the nucleic acid elution solution. Then lift the pipette upwards, remove the needle from the magnetic bead tube, and remove the magnetism.

[0041] S18, a) When the integrated reagent storage tube is set with the PCR amplification reagent tube, move the pipette above the PCR amplification reagent tube, move it downwards, and the needle enters the PCR amplification reagent tube. The air pump pushes air, which drives the pipette to flow the elution solution containing nucleic acid into the PCR amplification reagent tube. Control the air pump to push several times to mix the liquid. Then lift the pipette upwards and remove the needle from the PCR amplification reagent tube.

[0042] b) When a microfluidic chip is installed at the end of the connector of the integrated reagent storage tube, and PCR amplification reagents are pre-embedded in the microfluidic chip, move the pipette above the microfluidic chip, move it downward, the needle enters the microfluidic chip, the air pump pushes air, and the pipette drives the elution solution containing nucleic acid into the microfluidic chip.

[0043] c) When a microfluidic chip is installed at the end of the connector of the integrated reagent storage tube, but no PCR amplification reagent is pre-embedded in the microfluidic chip, move the pipette above the PCR amplification reagent tube, move it downwards, and the needle enters the PCR amplification reagent tube. The air pump pushes air, causing the pipette to flow the elution buffer containing nucleic acid into the PCR amplification reagent tube. Control the air pump to pump and push several times to mix the liquid. Then, lift the pipette upwards, and the needle is removed from the PCR amplification reagent tube. Then, move the pipette above the microfluidic chip, move it downwards, and the needle enters the microfluidic chip. The air pump pushes air, causing the pipette to bring the nucleic acid amplification detection reagent and the freshly extracted nucleic acid into the microfluidic chip.

[0044] S19. Set the temperature cycling program according to the requirements to perform temperature rise and fall and fluorescence signal acquisition on PCR amplification reagent tubes or microfluidic chips to achieve real-time fluorescence nucleic acid detection.

[0045] S20. After the reaction is complete, remove the entire integrated device, including the cartridge, pipette, and retractable elastic sealing cover, and discard it.

[0046] Compared with existing technologies, the sealed integrated device for nucleic acid detection using magnetic beads provided by this invention has the following advantages:

[0047] 1. Pre-filled reagents and automated mechanical operation for POCT testing: The integrated reagent storage tube of this invention arranges each reagent tube in a sequence strictly following manual operation steps. This arrangement, coordinated with the two-dimensional guide rails of the automated operating equipment, allows for convenient and rapid extraction and amplification of nucleic acid samples using magnetic beads, reducing manual operation and improving testing efficiency and accuracy. The cartridge of this invention seals the integrated reagent storage tube with a sealing gasket. Liquid transfer is performed using a needle pipette, and the retractable elastic sealing cover ensures the reagents remain sealed throughout the testing process, preventing contact with the external environment, reducing contamination, expanding the applicable scenarios for nucleic acid testing, and facilitating POCT nucleic acid testing.

[0048] 2. Wide range of applications and easy to upgrade to microfluidic chip detection: The integrated reagent storage tube of this invention has reserved space for microfluidic chips, which facilitates the overall upgrade of equipment and detection projects in the future. The cartridge and pipette of this invention have a wide range of applications and can be extended to all projects that currently require nucleic acid testing.

[0049] 3. Simple structure and easy to match with existing testing facilities: The sealed integrated device for nucleic acid detection using magnetic beads of the present invention has a simple structure, but it meets the requirements of sealing and preventing contamination in each step of nucleic acid extraction and amplification detection. The key positions of each reagent tube have the same shape and structure as the manual operation steps. Therefore, it can be matched with existing testing facilities, such as heating, magnetic attraction, fluorescence and other equipment, and can be quickly promoted and widely applied.

[0050] 4. Double-layer protection, excellent sealing: For samples with particularly high sealing requirements, pipettes with rubber sleeves can be used. This provides double protection—the rubber sleeve and a retractable elastic sealing cover—as the needle moves between reagent tubes, ensuring the sealing and safety of the detection process. Pipettes with rubber sleeves have three states during use:

[0051] a. Liquid aspiration: When the pipette is vertically inserted downwards through the circular opening 17 of the cartridge cover 1, the rubber sleeve 4 will first contact the chamfered edge of the circular opening and the upper surface of the sealing gasket 2. The rubber sleeve 4 will then be unable to descend further, and the needle 5, encased in the rubber sleeve, will continue to descend, piercing the rubber sleeve 4 and the sealing gasket 2 in sequence, and inserting into the corresponding test tube below. The air pump is then turned on to draw air outwards, causing the liquid to flow through the needle 5 into the pipette 6. Because there is a filter layer 7 between the adapter 8 and the liquid in the pipette, the liquid will not contaminate the fixed adapter, the subsequent tubing, or the air pump.

[0052] b. Lifting Up: When the pipette is lifted upwards, the needle 5 will lift up first, but the rubber sleeve 4 will remain pressed against the circular opening 17 of the top cover. The needle 5 will then pass upwards through the sealing gasket 2 and the bottom of the rubber sleeve 4, finally retracting completely into the rubber sleeve 4. Subsequently, the entire pipette will be lifted to a certain height away from the circular opening 17. The needle hole of the sealing gasket will automatically close to prevent contamination. At the same time, since the needle 5 is wrapped in the rubber sleeve 4, it also prevents the liquid inside the needle 5 from dripping out and causing contamination during pipetting.

[0053] c. Pipetting: Move the pipette above the next test tube and insert it vertically downwards. The sleeve 4 will first contact the chamfered edge of the circular opening 17 and the upper surface of the sealing gasket 2. Then the sleeve 4 can no longer descend. The needle 5, which is wrapped by the sleeve, will continue to descend, piercing the sleeve 4 and the sealing gasket 2 in turn, and being inserted into the test tube. The air pump blows air inwards, pushing the liquid in the pipette 6 into the test tube. Attached Figure Description

[0054] Figure 1 This is a schematic diagram of the integrated device of the present invention;

[0055] Figure 2 This is a schematic diagram of the structure of the card box of the present invention, including the top cover, sealing gasket, and integrated reagent storage tube;

[0056] Figure 3 This is a schematic diagram of the pipette (including a rubber sleeve) of the present invention;

[0057] Figure 4 This is a schematic diagram showing a split cover on the card holder of the present invention;

[0058] Figure 5 This is a schematic diagram of the shape and structure of the sealing gasket of the present invention;

[0059] Figure 6 This is a schematic diagram of the integrated reagent storage tube upgraded with a microfluidic chip according to the present invention;

[0060] Figure 7 This is a cross-sectional view of the cartridge, pipette, and retractable elastic sealing cover of the present invention after installation (with microfluidic chip).

[0061] In the diagram: 1. Top cover; 2. Sealing gasket; 3. Integrated reagent storage tube; 4. Rubber sleeve; 5. Needle; 6. Pipette; 7. Filter layer; 8. ; 9. Magnetic bead tube; 10. Lysis buffer tube; 11. Washing buffer tube 1; 12. Washing buffer tube 2 (first); 13. Washing buffer tube 2 (second); 14. Elution buffer tube; 15. PCR amplification reagent tube; 16. Microfluidic chip; 17. Circular opening; 18. Recessed area; 19. Retractable elastic sealing cover; 20. Upper interface; 21. Split cap; 22. Lower interface; 23. Main body Detailed Implementation

[0062] To enable those skilled in the art to better understand the present application, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present application, and not all embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative effort should fall within the scope of protection of the present application.

[0063] It should be noted that the terms "first," "second," etc., in the specification, claims, and accompanying drawings of this application are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that such data can be interchanged where appropriate for the embodiments of this application described herein. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion; for example, a process, method, system, product, or apparatus that comprises a series of steps or units is not necessarily limited to those steps or units explicitly listed, but may include other steps or units not explicitly listed or inherent to such processes, methods, products, or apparatus.

[0064] In this application, the terms "upper," "lower," "left," "right," "front," "rear," "top," "bottom," "inner," "outer," "middle," "vertical," "horizontal," "lateral," and "longitudinal" indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. These terms are primarily for the purpose of better describing this application and its embodiments, and are not intended to limit the indicated device, element, or component to having a specific orientation, or to be constructed and operated in a specific orientation.

[0065] Furthermore, in addition to indicating location or positional relationship, some of the aforementioned terms may also have other meanings. For example, the term "above" may also be used in some cases to indicate a certain dependency or connection relationship. Those skilled in the art can understand the specific meaning of these terms in this application based on the specific circumstances.

[0066] Furthermore, the terms "installation," "setup," "equipped with," "connection," "linking," and "socketing" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral structure; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium, or an internal connection between two devices, components, or parts. Those skilled in the art can understand the specific meaning of these terms in this application based on the specific circumstances.

[0067] It should be noted that, unless otherwise specified, the embodiments and features described in this application can be combined with each other. This application will now be described in detail with reference to the accompanying drawings and embodiments.

[0068] Example 1: No microfluidic chip installed

[0069] See Figures 1-5A sealed integrated device for nucleic acid detection using magnetic beads is disclosed. The integrated device includes a cartridge, a pipette, and a retractable elastic sealing cover 19. The cartridge includes a top cover 1, a sealing gasket 2, and an integrated reagent storage tube 3. The integrated reagent storage tube 3 is composed of seven test tubes whose openings are sequentially fixedly connected to a connector. The detection reagent is pre-loaded into different test tubes of the integrated reagent storage tube 3, and then covered with the sealing gasket 2 and the top cover 1 to achieve reagent sealing within the cartridge. A circular opening 17 is provided on the top cover 1 corresponding to the position of each test tube. The pipette consists of a needle 5, a pipette tube 6, and a filter layer 7. The rear end of the needle 5 is connected to the pipette tube 6, and the filter layer 7 is fixed to the rear end of the pipette tube 6. The lower part of the retractable elastic sealing cover 19 is detachably and sealed to the upper part of the cartridge, covering all the circular openings 17 within the retractable elastic sealing cover 19. The pipette is placed inside the retractable elastic sealing cover 19, wherein the rear part of the pipette tube 6 is sealed to the upper part of the retractable elastic sealing cover 19.

[0070] In this embodiment, the retractable elastic sealing cover 19 is composed of an upper interface 20, a lower interface 22, and a body 23. The body 23 is made of an airtight and watertight elastic material, which is rubber in this embodiment. The pipette is sealed to the retractable elastic sealing cover 19 through the upper interface 20. The upper part of the cartridge is connected to the retractable elastic sealing cover 19 through the lower interface 22. The body 23 is shaped like an accordion-style triangular cylinder.

[0071] In this embodiment, the connector end of the integrated reagent storage tube 3 is not provided with a microfluidic chip 16, but in other embodiments, a microfluidic chip 16 may be provided. If a microfluidic chip 16 is provided, the part of the microfluidic chip 16 used for fluorescence collection is placed outside the area sealed and covered by the retractable elastic sealing cover 19.

[0072] In this embodiment, the test tubes of the integrated reagent storage tube 3 are arranged in a straight line.

[0073] In this embodiment, the rear end of the pipette 6 is connected to the adapter 8 of the matching machine, and the connection method includes threading.

[0074] In this embodiment, the pipette further includes a rubber sleeve 4, which wraps around the outside of the needle 5. The top end of the rubber sleeve 4 is arc-shaped, corresponding to the circular opening 17, and the diameter of the top end of the rubber sleeve 4 is larger than the diameter of the circular opening 17.

[0075] In this embodiment, the sealing gasket 2 is set in the shape of a recess 18 corresponding to the position of each test tube opening.

[0076] In this embodiment, the material of the rubber sleeve 4 and the sealing gasket 2 is rubber, which is elastic and can be punctured by a needle.

[0077] In this embodiment, the test tubes of the integrated reagent storage tube 3 are, in sequence: magnetic bead tube 9, lysis buffer tube 10, washing buffer tube 11, washing buffer tube 2-12, washing buffer tube 2-2-13, elution buffer tube 14, and PCR amplification reagent tube 15.

[0078] In this embodiment, the cartridge, pipette, and retractable elastic sealing cover 19 are disposable.

[0079] In this embodiment, a separate cap 21 is provided on the card cover 1 of the card box at the position corresponding to the circular opening 17 of the lysis buffer tube 10, and the separate cap 21 penetrates the sealing gasket layer. Before testing, the separate cap can be opened to add the sample to be tested, and then the separate cap can be closed to keep it sealed. During the test, the needle of the pipette pierces through the separate cap and enters the test tube below for operation.

[0080] The detection method based on magnetic bead extraction and amplification for nucleic acid provided in this embodiment involves pre-adding the corresponding reagents to the integrated reagent storage tube 3, then adding the sample to the lysis buffer tube 10, sealing the retractable elastic sealing cover 19 and the pipette, and performing the detection in conjunction with automated operating equipment. The set operating steps are as follows:

[0081] S1. The adapter 8 is paired with an air pump. After connecting the adapter 8 to the pipette, it is moved above the lysis buffer tube 10. The pipette moves downward, and the rubber sleeve 4 rests against the circular opening 17. The needle 5 continues downward, piercing the rubber sleeve 4 and the sealing gasket 2 in sequence, and enters the lysis buffer tube 10. The air pump draws air, causing the pipette to draw all the reagents. Then the pipette is lifted upward, and the needle 5 is removed from the lysis buffer tube 10 and returned to the rubber sleeve 4. The sealing gasket 2 rebounds and seals the lysis buffer tube 10. The way the needle passes through the rubber sleeve to enter the reagent tube and removes the reagent tube to enter the rubber sleeve in the following steps is the same as in S1.

[0082] S2. Move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. The air pump pushes air, causing the pipette to inject all the liquid into the magnetic bead tube 9. Control the air pump to pump and push several times to mix the magnetic bead and liquid. Then lift the pipette upwards and remove the needle 5 from the magnetic bead tube 9.

[0083] S3. Heat the magnetic bead tube as needed. After stopping the heating, magnetically attract the magnetic bead tube to the side near the bottom for 1-5 minutes.

[0084] S4. Move the pipette downwards, the needle 5 enters the magnetic bead tube 9, the air pump draws air and drives the pipette to draw up all the reagents, then the pipette is lifted upwards, the needle 5 is removed from the magnetic bead tube 9, and the magnetism is removed.

[0085] S5. Move the pipette above the lysis buffer tube 10 and move it downwards. The needle 5 enters the lysis buffer tube 10. The air pump pushes air, which drives the pipette to inject the liquid into the lysis buffer tube 10. Then the pipette is lifted upwards and the needle 5 is removed from the lysis buffer tube 10.

[0086] S6. Move the pipette above the washing solution tube 11, move it downwards, and the needle 5 enters the washing solution tube 11. The air pump draws air to drive the pipette to draw all the reagents. Then the pipette is lifted upwards and the needle 5 is removed from the washing solution tube 11.

[0087] S7. Move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. The air pump pushes air to drive the pipette to inject the liquid into the magnetic bead tube 9. Control the air pump to pump and push several times to mix the magnetic bead with the liquid. Magnetic attraction is performed on the side of the magnetic bead tube near the bottom for 1-5 minutes. The air pump is then pumped to suck the waste liquid into the pipette. Then the pipette is lifted upwards, the needle 5 is removed from the magnetic bead tube 9, and the magnetic force is removed.

[0088] S8. Move the pipette above the washing liquid tube 11 and move it downwards. The needle 5 enters the washing liquid tube 11. The air pump pushes air, which drives the pipette to pump the waste liquid into the washing liquid tube 11. Then the pipette is lifted upwards and the needle 5 is removed from the washing liquid tube 11.

[0089] S9. Move the pipette above the washing solution tube 12 and move it downwards. The needle 5 enters the washing solution tube 12. The air pump draws air to drive the pipette to draw all the reagents. Then the pipette is lifted upwards and the needle 5 is removed from the washing solution tube 12.

[0090] S10, a) When the washing solution contains alcohol, move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. The air pump pushes air to drive the pipette to inject the liquid into the magnetic bead tube 9. Control the air pump to pump and push several times to mix the magnetic bead with the liquid. Magnetic attraction is performed on the side of the magnetic bead tube near the bottom for 1-5 minutes. The air pump is then pumped to suck the waste liquid into the pipette. Then the pipette is lifted upwards, the needle 5 is removed from the magnetic bead tube 9, and the magnetic force is removed.

[0091] b) When the washing solution does not contain alcohol, move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. The air pump pushes air, which drives the pipette to inject the liquid into the magnetic bead tube 9. Magnetic attraction is maintained on the side of the magnetic bead tube near the bottom for 1-5 minutes. Then the air pump is pumped out to suck the waste liquid into the pipette. Then the pipette is lifted upwards and the needle 5 is removed from the magnetic bead tube 9, maintaining the magnetic force.

[0092] S11. Move the pipette above the first 12 of the second washing liquid tube and move it downward. The needle 5 enters the first 12 of the second washing liquid tube. The air pump pushes air to drive the pipette to inject the waste liquid into the first 12 of the second washing liquid tube. Then the pipette is lifted upward and the needle 5 is removed from the first 12 of the second washing liquid tube.

[0093] S12. Move the pipette above the second washing solution tube 13 and move it downwards. The needle 5 enters the second washing solution tube 13. The air pump draws air to drive the pipette to draw up all the reagents. Then the pipette is lifted upwards and the needle 5 is removed from the second washing solution tube 13.

[0094] S13, a) When the washing solution contains alcohol, move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. The air pump pushes air, driving the pipette to inject the liquid into the magnetic bead tube 9. Control the air pump to pump and push several times to mix the magnetic bead with the liquid. Magnetic attraction is performed on the side of the magnetic bead tube near the bottom for 1-5 minutes. The air pump is then pumped to suck the waste liquid into the pipette. Then the pipette is lifted upwards, the needle 5 is removed from the magnetic bead tube 9, and the magnetic force is removed.

[0095] b) When the washing solution does not contain alcohol, move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. The air pump pushes air, which drives the pipette to inject the liquid into the magnetic bead tube 9. Then the air pump draws air to suck the waste liquid into the pipette. Then the pipette is lifted upwards, the needle 5 is removed from the magnetic bead tube 9, and the magnetism is removed.

[0096] S14. Move the pipette above the second washing liquid tube 13 and move it downward. The needle 5 enters the second washing liquid tube 13. The air pump pushes air to drive the pipette to flow the waste liquid into the second washing liquid tube 13. Then the pipette is lifted upward and the needle 5 is removed from the second washing liquid tube 13.

[0097] S15, a) When the washing solution contains alcohol, move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. Heat the magnetic bead tube 9 to 70°C for half a minute. Then lift the pipette upwards and remove the needle 5 from the magnetic bead tube 9.

[0098] b) This step is omitted if the second washing solution does not contain alcohol;

[0099] S16. Move the pipette above the eluent tube 14 and move it downwards. The needle 5 enters the eluent tube 14. The air pump draws air to drive the pipette to draw all the reagents. Then the pipette is lifted upwards and the needle 5 is removed from the eluent tube 14.

[0100] S17. Move the pipette above the magnetic bead tube 9 and move it downwards. Insert the needle 5 into the magnetic bead tube 9. Use the air pump to push air, which will cause the pipette to inject the elution solution into the magnetic bead tube 9. Control the air pump to pump and push several times to mix the magnetic bead with the liquid. Heat the magnetic bead tube 9 while heating. Control the air pump to pump and push several times. After heating, apply magnetic attraction to the side of the magnetic bead tube near the bottom for 1-5 minutes. Use the air pump to draw air and draw the elution solution containing nucleic acid into the pipette. Then lift the pipette upwards and remove the needle 5 from the magnetic bead tube 9 to remove the magnetic force.

[0101] S18. In this embodiment, the integrated reagent storage tube 3 is equipped with a PCR amplification reagent tube 15. The pipette is moved above the PCR amplification reagent tube 15 and then moved downwards. The needle 5 enters the PCR amplification reagent tube 15. The air pump pushes air, causing the pipette to flow the elution solution containing nucleic acid into the PCR amplification reagent tube 15. The air pump is controlled to pump and push several times to mix the liquid. Then the pipette is lifted upwards and the needle 5 is removed from the PCR amplification reagent tube 15.

[0102] S19. Set the temperature cycling program according to the requirements to perform temperature rise and fall and fluorescence signal acquisition on PCR amplification reagent tube 15 to achieve real-time fluorescence nucleic acid detection.

[0103] S20. After the reaction is complete, remove the entire integrated device, including the cartridge, pipette, and retractable elastic sealing cover 19, and dispose of it in the designated trash can.

[0104] Example 2: A microfluidic chip was used, but no PCR amplification reagents were pre-embedded.

[0105] See Figure 6 In this embodiment, a microfluidic chip 16 is provided at the end of the connector of the integrated reagent storage tube 3.

[0106] See Figure 7This embodiment provides a sealed integrated device for nucleic acid detection using magnetic beads. The integrated device includes a cartridge, a pipette, and a retractable elastic sealing cover 19. The cartridge includes a top cover 1, a sealing gasket 2, and an integrated reagent storage tube 3. The integrated reagent storage tube 3 is composed of seven test tubes whose openings are sequentially fixedly connected to a connector. The detection reagents are pre-filled into different test tubes of the integrated reagent storage tube 3, and then covered with the sealing gasket 2 and the top cover 1 to achieve a sealed reagent compartment. The top cover 1 has a corresponding label for each test tube. A circular opening 17 is provided at the position; the pipette consists of a needle 5, a pipette tube 6, and a filter layer 7; the rear end of the needle 5 is connected to the pipette tube 6, and the filter layer 7 is fixed to the rear end of the pipette tube 6; the lower part of the retractable elastic sealing cover 19 is detachably and sealed to the upper part of the cartridge, covering all the circular openings 17 in the retractable elastic sealing cover 19; the pipette is placed inside the retractable elastic sealing cover 19, wherein the rear part of the pipette tube 6 is sealed and connected to the upper part of the retractable elastic sealing cover 19.

[0107] In this embodiment, the retractable elastic sealing cover 19 is composed of an upper interface 20, a lower interface 22, and a body 23. The body 23 is made of airtight and watertight rubber. The pipette is sealed to the retractable elastic sealing cover 19 through the upper interface 20. The upper part of the cartridge is connected to the retractable elastic sealing cover 19 through the lower interface 22. The body 23 is shaped like an accordion-style triangular cylinder.

[0108] In this embodiment, a microfluidic chip 16 is provided at the end of the connector of the integrated reagent storage tube 3.

[0109] In this embodiment, the portion of the microfluidic chip 16 used for fluorescence acquisition is placed outside the area sealed and covered by the retractable elastic sealing cover 19.

[0110] In this embodiment, the test tubes of the integrated reagent storage tube 3 are arranged in a straight line.

[0111] In this embodiment, the rear end of the pipette 6 is connected to the adapter 8 of the matching machine, and the connection method is threaded.

[0112] In this embodiment, the sealing gasket 2 is flat.

[0113] In this embodiment, the rubber sleeve 4 and the sealing gasket 2 are made of elastic rubber that can be punctured by a needle.

[0114] In this embodiment, the test tubes of the integrated reagent storage tube 3 are, in sequence: magnetic bead tube 9, lysis buffer tube 10, washing buffer tube 11, washing buffer tube 2-12, washing buffer tube 2-2-13, elution buffer tube 14, and PCR amplification reagent tube 15.

[0115] In this embodiment, the microfluidic chip 16 does not have PCR amplification reagents pre-embedded.

[0116] In this embodiment, the cartridge and pipette are disposable.

[0117] This invention also provides a detection method based on magnetic bead-based nucleic acid extraction and amplification. In this method, the corresponding reagents are first added to the integrated reagent storage tube 3, then the sample is added to the lysis buffer tube 10. A retractable elastic sealing cover 19 and a pipette are then connected and installed. The lower interface of the retractable elastic sealing cover 19 with the pipette installed is then sealed to the lower part of the cartridge. After installation, detection is performed using automated operating equipment. The set operating steps are as follows:

[0118] S1. The adapter 8 is equipped with an air pump. After connecting the adapter 8 to the pipette, it is moved above the lysis buffer tube 10. The pipette moves downward, and the needle 5 continues downward, piercing the sealing gasket 2 and entering the lysis buffer tube 10. The air pump draws air to drive the pipette to draw all the reagents. Then the pipette is lifted upward, and the needle 5 is removed from the lysis buffer tube 10. The sealing gasket 2 springs back to seal the lysis buffer tube 10.

[0119] S2. Move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. The air pump pushes air, causing the pipette to inject all the liquid into the magnetic bead tube 9. Control the air pump to pump and push several times to mix the magnetic bead and liquid. Then lift the pipette upwards and remove the needle 5 from the magnetic bead tube 9.

[0120] S3. Heat the magnetic bead tube as needed. After stopping the heating, magnetically attract the magnetic bead tube to the side near the bottom for 1-5 minutes.

[0121] S4. Move the pipette downwards, the needle 5 enters the magnetic bead tube 9, the air pump draws air and drives the pipette to draw up all the reagents, then the pipette is lifted upwards, the needle 5 is removed from the magnetic bead tube 9, and the magnetism is removed.

[0122] S5. Move the pipette above the lysis buffer tube 10 and move it downwards. The needle 5 enters the lysis buffer tube 10. The air pump pushes air, which drives the pipette to inject the liquid into the lysis buffer tube 10. Then the pipette is lifted upwards and the needle 5 is removed from the lysis buffer tube 10.

[0123] S6. Move the pipette above the washing solution tube 11, move it downwards, and the needle 5 enters the washing solution tube 11. The air pump draws air to drive the pipette to draw all the reagents. Then the pipette is lifted upwards and the needle 5 is removed from the washing solution tube 11.

[0124] S7. Move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. The air pump pushes air to drive the pipette to inject the liquid into the magnetic bead tube 9. Control the air pump to pump and push several times to mix the magnetic bead with the liquid. Magnetic attraction is performed on the side of the magnetic bead tube near the bottom for 1-5 minutes. The air pump is then pumped to suck the waste liquid into the pipette. Then the pipette is lifted upwards, the needle 5 is removed from the magnetic bead tube 9, and the magnetic force is removed.

[0125] S8. Move the pipette above the washing liquid tube 11 and move it downwards. The needle 5 enters the washing liquid tube 11. The air pump pushes air, which drives the pipette to pump the waste liquid into the washing liquid tube 11. Then the pipette is lifted upwards and the needle 5 is removed from the washing liquid tube 11.

[0126] S9. Move the pipette above the washing solution tube 12 and move it downwards. The needle 5 enters the washing solution tube 12. The air pump draws air to drive the pipette to draw all the reagents. Then the pipette is lifted upwards and the needle 5 is removed from the washing solution tube 12.

[0127] S10, a) When the washing solution contains alcohol, move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. The air pump pushes air to drive the pipette to inject the liquid into the magnetic bead tube 9. Control the air pump to pump and push several times to mix the magnetic bead with the liquid. Magnetic attraction is performed on the side of the magnetic bead tube near the bottom for 1-5 minutes. The air pump is then pumped to suck the waste liquid into the pipette. Then the pipette is lifted upwards, the needle 5 is removed from the magnetic bead tube 9, and the magnetic force is removed.

[0128] b) When the washing solution does not contain alcohol, move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. The air pump pushes air, which drives the pipette to inject the liquid into the magnetic bead tube 9. Magnetic attraction is maintained on the side of the magnetic bead tube near the bottom for 1-5 minutes. Then the air pump is pumped out to suck the waste liquid into the pipette. Then the pipette is lifted upwards and the needle 5 is removed from the magnetic bead tube 9, maintaining the magnetic force.

[0129] S11. Move the pipette above the first 12 of the second washing liquid tube and move it downward. The needle 5 enters the first 12 of the second washing liquid tube. The air pump pushes air to drive the pipette to inject the waste liquid into the first 12 of the second washing liquid tube. Then the pipette is lifted upward and the needle 5 is removed from the first 12 of the second washing liquid tube.

[0130] S12. Move the pipette above the second washing solution tube 13 and move it downwards. The needle 5 enters the second washing solution tube 13. The air pump draws air to drive the pipette to draw up all the reagents. Then the pipette is lifted upwards and the needle 5 is removed from the second washing solution tube 13.

[0131] S13, a) When the washing solution contains alcohol, move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. The air pump pushes air, driving the pipette to inject the liquid into the magnetic bead tube 9. Control the air pump to pump and push several times to mix the magnetic bead with the liquid. Magnetic attraction is performed on the side of the magnetic bead tube near the bottom for 1-5 minutes. The air pump is then pumped to suck the waste liquid into the pipette. Then the pipette is lifted upwards, the needle 5 is removed from the magnetic bead tube 9, and the magnetic force is removed.

[0132] b) When the washing solution does not contain alcohol, move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. The air pump pushes air, which drives the pipette to inject the liquid into the magnetic bead tube 9. Then the air pump draws air to suck the waste liquid into the pipette. Then the pipette is lifted upwards, the needle 5 is removed from the magnetic bead tube 9, and the magnetism is removed.

[0133] S14. Move the pipette above the second washing liquid tube 13 and move it downward. The needle 5 enters the second washing liquid tube 13. The air pump pushes air to drive the pipette to flow the waste liquid into the second washing liquid tube 13. Then the pipette is lifted upward and the needle 5 is removed from the second washing liquid tube 13.

[0134] S15, a) When the washing solution contains alcohol, move the pipette above the magnetic bead tube 9 and move it downwards. The needle 5 enters the magnetic bead tube 9. Heat the magnetic bead tube 9 to 70°C for half a minute. Then lift the pipette upwards and remove the needle 5 from the magnetic bead tube 9.

[0135] b) This step is omitted if the second washing solution does not contain alcohol;

[0136] S16. Move the pipette above the eluent tube 14 and move it downwards. The needle 5 enters the eluent tube 14. The air pump draws air to drive the pipette to draw all the reagents. Then the pipette is lifted upwards and the needle 5 is removed from the eluent tube 14.

[0137] S17. Move the pipette above the magnetic bead tube 9 and move it downwards. Insert the needle 5 into the magnetic bead tube 9. Use the air pump to push air, which will cause the pipette to inject the elution solution into the magnetic bead tube 9. Control the air pump to pump and push several times to mix the magnetic bead with the liquid. Heat the magnetic bead tube 9 while heating. Control the air pump to pump and push several times. After heating, apply magnetic attraction to the side of the magnetic bead tube near the bottom for 1-5 minutes. Use the air pump to draw air and draw the elution solution containing nucleic acid into the pipette. Then lift the pipette upwards and remove the needle 5 from the magnetic bead tube 9 to remove the magnetic force.

[0138] S18. Move the pipette above the PCR amplification reagent tube 15 and move it downwards. The needle 5 enters the PCR amplification reagent tube 15. The air pump pushes air, causing the pipette to flow the elution solution containing nucleic acid into the PCR amplification reagent tube 15. Control the air pump to push several times to mix the liquid. Then lift the pipette upwards and remove the needle 5 from the PCR amplification reagent tube 15.

[0139] S19. Move the pipette above the microfluidic chip 16 and downwards. The needle 5 enters the inlet of the microfluidic chip 16. The air pump pushes air, causing the pipette to flow the mixture of nucleic acid amplification reagent and extracted nucleic acid into the microfluidic chip 16. Then, the pipette is lifted upwards, and the needle 5 is removed from the microfluidic chip 16. Set the temperature cycling program as needed to perform temperature cycling amplification on the microfluidic chip 16 and collect fluorescence signals to achieve real-time fluorescence nucleic acid detection.

[0140] S20. After the reaction is complete, remove the entire integrated device, including the cartridge, pipette, and retractable elastic sealing cover 19, and dispose of it in the designated trash can.

[0141] The integrated device of this invention can be matched with supporting automated testing equipment to achieve rapid and efficient testing. Since the entire operation is sealed, the sample will not leak or be contaminated. Therefore, there are no requirements for the laboratory and it can be operated in a general laboratory to meet the needs of POCT.

[0142] Using the detection device of this invention, operated on compatible automated equipment, results for COVID-19 nucleic acid testing can be obtained within one hour. Even without such automated equipment, personnel can manually operate the device using a handheld pipette on the cartridge, also obtaining results within one hour. In contrast, in a conventional PCR laboratory, due to the need for transfer between different laboratory areas, results typically take 2-3 hours under ideal conditions. Therefore, practical experience demonstrates that the sealed, integrated device for nucleic acid extraction and amplification using magnetic beads of this invention can meet the needs of point-of-care testing (POCT) and has promising market application prospects.

[0143] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A sealed integrated device for magnetic bead method nucleic acid detection, characterized by, The integrated device comprises a cartridge, a pipette and a telescopic elastic seal cover (19), the cartridge comprises an upper cover (1), a sealing gasket (2) and an integrated reagent storage tube (3), the integrated reagent storage tube (3) is composed of a plurality of open parts of test tubes which are sequentially fixed and connected on a connecting body, the detection reagent is preloaded into different test tubes of the integrated reagent storage tube (3), then the sealing gasket (2) and the upper cover (1) are covered to realize the sealing of the reagent in the cartridge; a circular opening (17) is arranged on the upper cover (1) at a position corresponding to each test tube; the pipette is composed of a needle (5), a pipette (6) and a filter layer (7); the rear end of the needle (5) is connected with the pipette (6), and the filter layer (7) is fixed at the rear end of the pipette (6); the lower part of the telescopic elastic seal cover (19) is detachably and sealingly connected with the upper part of the cartridge, and all the circular openings (17) are covered in the telescopic elastic seal cover (19); the pipette is arranged in the telescopic elastic seal cover (19), wherein the rear part of the pipette (6) is sealingly connected with the upper part of the telescopic elastic seal cover (19). The telescopic elastic seal cover (19) is composed of an upper interface (20), a lower interface (22) and a body (23), the body (23) is made of an elastic material which is air and water impermeable; the pipette is sealingly connected with the telescopic elastic seal cover (19) through the upper interface (20); the upper part of the cartridge is connected with the telescopic elastic seal cover (19) through the lower interface (22); the shape of the body (23) comprises a flat plate type triangular cylinder, a folded triangular cylinder and an organ type triangular cylinder. A microfluidic chip (16) is arranged at the end of the connecting body of the integrated reagent storage tube (3); The part of the microfluidic chip (16) for fluorescence collection is arranged outside the area covered by the telescopic elastic seal cover (19); The pipette further comprises a rubber sleeve (4), the rubber sleeve (4) is wrapped outside the needle (5), the top end of the rubber sleeve (4) is in a circular arc shape, corresponds to the circular opening (17) and has a diameter greater than that of the circular opening (17); The test tubes of the integrated reagent storage tube (3) are sequentially a magnetic bead tube (9), a lysis solution tube (10), a washing solution one tube (11), a washing solution two tube one (12), a washing solution two tube two (13), an eluent tube (14) and a PCR amplification reagent tube (15); The sealing gasket (2) is recessed (18) at a position corresponding to the opening of each test tube; a split cover (21) is arranged on the upper cover (1) of the cartridge at a position corresponding to the circular opening (17) of the lysis solution tube (10), and the split cover (21) penetrates the sealing gasket layer.

2. The sealed integrated device for magnetic bead-based nucleic acid detection according to claim 1, wherein The plurality of test tubes of the integrated reagent storage tube (3) are arranged in a straight line, a circle or an array.

3. The sealed integrated device for magnetic bead-based nucleic acid detection according to claim 1, wherein The rear end of the pipette (6) is connected with an adapter (8) of a matching machine in a connection mode including screw thread, buckle and tight fit.

4. The sealed integrated device for magnetic bead-based nucleic acid detection according to claim 1, wherein The materials of the rubber sleeve (4) and the sealing gasket (2) are materials which are elastic and can be pierced by the needle, including rubber and silicone.

5. The sealed integrated device for magnetic bead-based nucleic acid detection according to claim 1, wherein The cartridge, pipette and retractable, flexible seal (19) are disposable.

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