A nucleic acid detection kit

Abstract

The present invention discloses a nucleic acid detection kit, comprising an extraction chip, an amplification chip and a control valve packaged into one body, wherein the extraction chip comprises a plurality of chambers arranged circumferentially and a rotary valve arranged rotatably, wherein some of the chambers are lysis and binding chambers, cleaning reaction chambers and elution reaction chambers, and the rotary valve has an extraction port, which is connected to one of the chambers as the rotary valve rotates; the amplification chip is stacked with the extraction chip, and the amplification chip has an amplification reaction chamber, and the control valve is used to control the connection or disconnection between the elution reaction chamber and the amplification reaction chamber, so that the object to be detected is transferred to the amplification reaction chamber. The nucleic acid detection kit provided by the present invention, in conjunction with an external control instrument, realizes fully automatic detection of nucleic acids in a closed space, improves the accuracy and stability of nucleic acid detection, and can avoid various problems such as reagent contamination and environmental pollution during nucleic acid detection.

Description

Technical Field

[0001] The present invention relates to the technical field of medical devices, and in particular to a nucleic acid detection kit and a control method thereof. Background Art

[0002] Molecular diagnosis refers to the use of molecular biology methods to detect changes in the structure or expression levels of a patient's genetic material. Molecular diagnosis is a primary method for predictive diagnosis, enabling both diagnosis of individual genetic diseases and prenatal diagnosis. Molecular diagnosis primarily involves the detection of genes encoding disease-related structural proteins, enzymes, antigens, antibodies, and immunoreactive molecules. Key molecular diagnostic techniques include nucleic acid hybridization, polymerase chain reaction, and biochip technology.

[0003] Among them, PCR (polymerase chain reaction) products occupy the main market share of molecular diagnostics. PCR, a full-process polymerase chain reaction, utilizes the fact that DNA denatures into single strands at a high temperature of 95°C in vitro. At low temperatures (often around 60°C), primers bind to the single strands according to the principle of base complementary pairing. The temperature is then adjusted to the optimal reaction temperature of DNA polymerase (around 72°C), and DNA polymerase synthesizes complementary chains along the direction from phosphate to pentose (5'-3'). PCR products have high sensitivity, strong specificity, and a short diagnostic window period. They can perform qualitative and quantitative testing and are widely used for hepatitis, sexually transmitted diseases, lung infectious diseases, eugenics, genetic diseases, tumors, etc., filling the detection gap in the early immune detection window period and providing effective assistance for early diagnosis, early treatment, and safe blood use.

[0004] Nucleic acid testing of samples generally involves three steps: nucleic acid extraction, nucleic acid amplification, and nucleic acid detection. Currently, commercial nucleic acid testing products mostly perform nucleic acid extraction, nucleic acid amplification, and nucleic acid detection independently. After the preceding steps are completed, the sample is moved to the subsequent equipment for completion. Consequently, the preceding steps cannot be effectively integrated with the subsequent steps for continuous execution.

[0005] Existing nucleic acid detection reagent products that complete each step independently require independent equipment to complete each step. Multiple devices are needed in a single nucleic acid detection process, and the equipment takes up a large space. Furthermore, after the previous step is completed, the sample needs to be moved to the subsequent equipment, which is cumbersome and time-consuming, and has high requirements on the environment and personnel. At the same time, for non-integrated reagent products, when switching from the previous step to the subsequent step, the sample is also susceptible to contamination from the external environment or contamination of the detection environment during the movement process.

[0006] Microfluidic chip technology utilizes highly integrated functional units to implement multi-step biochemical reactions at a microscale, reducing manual operations and enabling automated sample-in, result-out solutions. In recent years, it has attracted widespread attention and applied research in fields such as biomedical diagnostics, analytical chemistry, and life sciences. The advantages of microfluidic chips include reduced reaction volumes and reagent requirements, significantly shortening sample processing time, a fully enclosed chip-reagent integration system that prevents cross-contamination, and automated control that reduces manual operations and improves operational consistency.

[0007] Therefore, it is necessary to provide a nucleic acid detection kit that uses microfluidic chip technology to achieve fully enclosed integrated nucleic acid extraction and detection processing. Summary of the Invention

[0008] The purpose of the present invention is to provide a nucleic acid detection kit that can realize fully enclosed integrated processing of nucleic acid extraction, amplification and detection.

[0009] To achieve the above-mentioned purpose, the technical solution adopted by the present invention is to provide a nucleic acid detection kit, including an extraction chip, an amplification chip and a control valve packaged into one body, the extraction chip including multiple chambers arranged in a circumferential direction and a rotary valve arranged in a rotational direction, wherein some chambers are lysis and binding chambers, cleaning reaction chambers and elution reaction chambers, the rotary valve has an extraction port, and the extraction port is connected to one of the chambers as the rotary valve rotates; the amplification chip is stacked with the extraction chip, the amplification chip has an amplification reaction chamber, and the control valve is used to control the connection or disconnection of the elution reaction chamber and the amplification reaction chamber, so that the object to be detected is transferred to the amplification reaction chamber.

[0010] Preferably, it includes a top cover, which is arranged on the extraction chip, and is provided with multiple sampling holes, multiple ventilation holes and a through hole for providing pressure. The multiple sampling holes are respectively connected to the cleavage and combination chamber, the cleaning reaction chamber and the elution reaction chamber, the multiple ventilation holes are respectively connected to the cleavage and combination chamber and the cleaning reaction chamber, and the through hole for providing pressure is connected to the elution reaction chamber through a microchannel.

[0011] Preferably, a plurality of first scale lines are provided on the top cover, a second scale line is provided corresponding to the rotary valve, and a third scale line is provided corresponding to the control valve.

[0012] Preferably, the rotary valve includes a first rotating shaft and a first sealing gasket, the first sealing gasket is sleeved on one end of the first rotating shaft, the extraction port is provided on the first sealing gasket, and the second scale line is provided on the other end of the first rotating shaft.

[0013] Preferably, the first rotating shaft has a cavity for accommodating the magnet, a flange is provided on the outside of the first rotating shaft, the inner side surface of the first sealing gasket is in contact with the first rotating shaft, and the end surface of the first sealing gasket is in contact with the flange.

[0014] Preferably, a lysis and combination chamber, a first cleaning reaction chamber, a second cleaning reaction chamber, a first drying chamber, an elution reaction chamber and a second drying chamber are sequentially arranged on the extraction chip along the rotation direction of the rotary valve. The second drying chamber isolates the elution reaction chamber and the lysis and combination chamber. The first drying chamber and the second drying chamber are both connected to a through hole that provides pressure.

[0015] Preferably, the cleavage and combination chamber, the first cleaning reaction chamber, the second cleaning reaction chamber, and the elution reaction chamber are arranged from the top surface to the bottom surface of the extraction chip, and the first drying chamber and the second drying chamber are arranged on the bottom surface of the extraction chip and do not pass through the extraction chip.

[0016] Preferably, the control valve includes a second rotating shaft, the third scale line is set on the side of one end of the second rotating shaft, and a microchannel is set on the end face of the other end of the second rotating shaft. The direction of the microchannel on the control valve is controlled by rotating the second rotating shaft to connect or disconnect the elution reaction chamber and the amplification reaction chamber.

[0017] Preferably, it includes a fixing ring and a second sealing gasket, the extraction chip has a first groove, the control valve is arranged in the first groove, the fixing ring and the second sealing gasket are matched and assembled, and the control valve is fixed in the fixing ring; the second sealing gasket is connected to the bottom of the first groove.

[0018] Preferably, the amplification chip is provided with four amplification reaction chambers for placing different amplification reagents, and the four amplification reaction chambers are connected through a microfluidic channel.

[0019] Preferably, each of the amplification reaction chambers is provided with a vent hole, and a waterproof breathable membrane is provided corresponding to the vent hole.

[0020] Preferably, it comprises a first sealing film and a second sealing film, wherein the first sealing film is arranged between the extraction chip and the amplification chip, and the bottom surface of the amplification chip is attached to the second sealing film.

[0021] Compared with the prior art, the present invention has the following beneficial effects: the nucleic acid detection kit provided by the present invention has the following advantages:

[0022] 1. By packaging the extraction chip, amplification chip, rotary valve, and control valve into one, nucleic acid detection reagents can be integrated into a microfluidic chip; the kit has a compact structure, small size, easy to carry, flexible use, and can adapt to a variety of environments.

[0023] 2. The extraction chip is sealed by the top cover and the first sealing film, and the amplification chip is sealed by the first sealing film and the second sealing film. The entire process of nucleic acid detection, nucleic acid extraction, purification, amplification, detection, etc. can be carried out in a closed space.

[0024] 3. By setting up multiple amplification reaction chambers on the amplification chip and encapsulating different freeze-dried reaction reagents, multiple nucleic acid detection is achieved in one chip system, which improves the nucleic acid detection throughput and reduces the amount of detection samples.

[0025] 4. The nucleic acid detection kit provided by the present invention cooperates with an external control instrument to realize fully automatic nucleic acid detection. All processes of nucleic acid detection are completed automatically in a relatively short time, with fewer operating steps, which reduces personnel time and reduces the errors and uncertainties caused by manual operation to nucleic acid detection, thereby improving the accuracy and stability of nucleic acid detection. At the same time, it can avoid various problems such as reagent contamination and environmental pollution during the nucleic acid detection process. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 A schematic diagram of an overall explosion of the nucleic acid detection kit provided by an embodiment of the present invention;

[0027] Figure 2a The top view of the chip is extracted in the embodiment of the present invention. Figure 2b A bottom view of a chip is extracted in an embodiment of the present invention;

[0028] Figure 3 Schematic diagram of the structure of a rotary valve in an embodiment of the present invention;

[0029] Figure 4 This is a schematic diagram of the structure of the amplification chip in an embodiment of the present invention;

[0030] Figure 5 This is a schematic structural diagram of a top cover in an embodiment of the present invention;

[0031] Figure 6 This is a schematic structural diagram of a control valve in an embodiment of the present invention;

[0032] Figure 7 Schematic diagram of the structure of the first sealing film in an embodiment of the present invention;

[0033] Figure 8 Schematic diagram of the structure of the second sealing film in an embodiment of the present invention;

[0034] Figure 9 This is a schematic structural diagram of a nucleic acid detection kit according to an embodiment of the present invention;

[0035] Figure 10 This is a schematic diagram of the initial state of the chip after packaging in an embodiment of the present invention;

[0036] Figure 11 Schematic diagram of a nucleic acid detection kit according to an embodiment of the present invention.

[0037] In the picture:

[0038] 1-top cover, 2-control valve, 3-fixing ring, 4-rotary valve, 5-second sealing gasket, 6-extraction chip, 7-amplification chip, 8-first sealing membrane, 9-second sealing membrane, 1001, 1002, 1003, 1015-injection hole, 1004, 1005, 1006, 1007-through hole, 1008, 1009, 1010-vent, 1011, 1012, 1013, 1014-first scale line, 2001-second rotating shaft, 2002-third scale line, 2003-microfluidic channel, 2004-second flange, 4001-first rotating shaft, 4002-first sealing gasket, 40011-second scale line, 40021-extraction port, 6001-lysis and binding chamber, 6002-first cleaning reaction Chamber, 6003-second cleaning reaction chamber, 6004-mounting hole, 6005, 6012-through hole, 6006, 6011, 6015, 6016, 6017-microchannel, 6007-first groove, 6008, 6009-through hole, 6010-elution reaction chamber, 6013-first drying chamber, 6014-second drying chamber, 7001, 70 02, 7003, 7004, 7005-through hole, 7006-microfluidic channel, 7007, 7008, 7009, 7010-amplification reaction chamber, 7011, 7012, 7013, 7014-vents, 8001-through hole, 8002, 8003, 8004, 8005-vents, 9001, 9002, 9003-through hole. DETAILED DESCRIPTION

[0039] The present invention will be further described below with reference to the accompanying drawings and examples.

[0040] It should be noted that the terms "inside," "outside," "upper," "lower," and similar expressions used in this invention are for illustrative purposes only and do not represent the only implementation methods. The terms "installed," "connected," and "connected" should be understood broadly. For example, they can refer to fixed connections, removable connections, or integral connections; they can be directly connected or indirectly connected through an intermediate medium, or they can be internal connections between two components. The term "microchannel" refers to a long, narrow slit. Those skilled in the art will understand the specific meanings of these terms in this invention based on specific circumstances.

[0041] See Figure 1 、 Figure 2a and Figure 4The nucleic acid detection kit provided by the present invention includes an extraction chip 6, an amplification chip 7, and a control valve 2, which are packaged into one body. The extraction chip 6 includes a plurality of chambers arranged circumferentially and a rotary valve 4 arranged rotatably, wherein some of the chambers are a lysis and binding chamber 6001, a cleaning reaction chamber, and an elution reaction chamber 6010. The rotary valve 4 has an extraction port 40021. As the rotary valve 4 rotates, the extraction port 40021 communicates with one of the chambers and is used to control the transfer of the object to be detected from the lysis and binding chamber 6001 to the cleaning reaction chamber and the elution reaction chamber 6010 in sequence. The amplification chip 7 is stacked with the extraction chip 6, and the amplification chip 7 has an amplification reaction chamber 7007. The control valve 2 is used to control the connection or disconnection between the elution reaction chamber and the amplification reaction chamber, so that the object to be detected is transferred to the amplification reaction chamber. Furthermore, there are two cleaning reaction chambers, namely a first cleaning reaction chamber 6002 and a second cleaning reaction chamber 6003.

[0042] See Figure 2a and Figure 2bIn a specific embodiment, a lysis and binding chamber 6001, a first cleaning reaction chamber 6002, a second cleaning reaction chamber 6003, a first drying chamber 6013, an elution reaction chamber 6010, and a second drying chamber 6014 are sequentially arranged on the extraction chip 6 along the rotation direction of the rotary valve 4. During detection, a lysis solution containing magnetic beads is encapsulated in the lysis and binding chamber 6001. The rotary valve 4 rotates and combines with the magnetic attraction of an external device to transfer the magnetic beads bound to the nucleic acid from the lysis and binding chamber 6001 to the first cleaning reaction chamber 6002, the second cleaning reaction chamber 6003, the first drying chamber 6013, and the elution reaction chamber 6010 in sequence. The second drying chamber 6014 isolates the elution reaction chamber 6010 from the lysis and binding chamber 6001 to prevent solutions from other chambers from leaking into the elution reaction chamber 6010 and affecting the detection results. Furthermore, the lysis and binding chamber 6001, the first cleaning reaction chamber 6002, the second cleaning reaction chamber 6003, and the elution reaction chamber 6010 extend from the top to the bottom of the extraction chip 6. The first drying chamber 6013 and the second drying chamber 6014 are located on the bottom of the extraction chip 6 but do not penetrate the chip 6. The lysis and binding chamber 6001, the first cleaning reaction chamber 6002, the second cleaning reaction chamber 6003, and the elution reaction chamber 6010 serve as the reaction chambers for the entire nucleic acid extraction and purification process. The first drying chamber 6013 and the second drying chamber 6014 are located on either side of the elution reaction chamber 6010. The first drying chamber 6013 is primarily used for drying the magnetic beads before elution and is connected to the through-hole 6005 via a microfluidic channel 6016. The accompanying instrument uses a piston to repeatedly apply positive and negative pressure from the through-hole 6005 to the first drying chamber 6013, effectively drying the magnetic beads. The second drying chamber 6014 is primarily used to isolate the elution reaction chamber 6010 from the other reaction chambers. The second drying chamber 6014 is connected to the through hole 6012 via the microchannel 6015 ; the piston on the supporting instrument can apply a certain pressure to the elution reaction chamber 6010 through the through hole 6012 .

[0043] The extraction chip 6 is also provided with a mounting hole 6004 and a first groove 6007. The first groove 6007 is preferably a circular groove. The control valve 2 is installed in the first groove 6007, and the rotary valve 4 is installed in the mounting hole 6004. One end of the microchannel 6017 is connected to the elution reaction chamber 6010, and the other end extends to the control valve 2 for communication with the microchannel 2003 on the control valve 2. During use, by rotating the control valve 2, the eluate after the reaction can pass through the microchannel 6017 and the microchannel 2003 and enter the amplification chip 7 for subsequent amplification reaction.

[0044] See Figure 3The rotary valve 4 includes a first rotating shaft 4001 and a first sealing gasket 4002. The first end of the rotating shaft 4001 is tightly covered with the first sealing gasket 4002. The first sealing gasket 4002 is provided with an extraction port 40021. The magnetic beads bound to the nucleic acid can be adsorbed into the extraction port 40021. The outer side of the second end of the first rotating shaft 4001 is provided with a second scale line 40011. The second scale line 40011 cooperates with the first scale lines 1011, 1012, and 1013 provided on the top cover 1 to display the rotation angle of the rotary valve 4, so that the extraction port 40021 is completely aligned with the lysis and combination chamber 6001, the first cleaning reaction chamber 6002 or the second cleaning reaction chamber 6003. At the same time, the first sealing gasket 4002 seals each chamber to prevent solution leakage. Furthermore, the first rotating shaft 4001 has a cavity for accommodating a magnet. A magnet inserted into the cavity moves the magnetic beads toward the center and secures them there. The first rotating shaft 4001 and the first sealing gasket 4002 are tightly bonded to prevent leakage and seepage. Furthermore, the first sealing gasket 4002 is made of a soft material, such as silicone, for excellent sealing performance. The lysis and binding chamber 6001, the first cleaning reaction chamber 6002, the second cleaning reaction chamber 6003, the first drying chamber 6013, the elution reaction chamber 6010, and the second drying chamber 6014 are evenly distributed around the rotary valve 4. Every 60° rotation of the rotary valve 4 aligns the extraction port 40021 with the next chamber.

[0045] See Figure 4In one embodiment, the amplification chip 7 is provided with four amplification reaction chambers 7007, 7008, 7009, and 7010 for storing different fixed volumes of freeze-dried amplification reagents. The four amplification reaction chambers 7007, 7008, 7009, and 7010 are connected by a microfluidic channel 7006. The four amplification reaction chambers 7007, 7008, 7009, and 7010 are respectively provided with vents 7011, 7012, 7013, and 7014. The vents 7011, 7012, 7013, and 7014 are respectively provided with waterproof and breathable membranes to ensure that air is discharged while the eluted reaction solution fills the four amplification reaction chambers 7007, 7008, 7009, and 7010 and the microfluidic channel 7006. Furthermore, the amplification chip 7 is provided with four through holes 7001, 7002, 7003, and 7004. The shapes and positions of through holes 7001, 7002, and 7003 correspond to the lysis and binding chamber 6001, the first cleaning reaction chamber 6002, and the second cleaning reaction chamber 6003, respectively. During operation, an ultrasonic head can be inserted into through holes 7001, 7002, and 7003 to ultrasonically vibrate the reaction solutions in the lysis and binding chamber 6001, the first cleaning reaction chamber 6002, and the second cleaning reaction chamber 6003. The shape and position of through hole 7004 correspond to the mounting hole 6004. During operation, a permanent magnetic head can be inserted into through hole 7004 to attract magnetic beads to the extraction port 40021. The amplification chip 7 is bonded to the extraction chip 6 by screws, hot pressing, laser, or ultrasonic welding. The amplification chip 7 is also provided with a through hole 7005 connected to the microchannel 7006. The through hole 7005 and the microchannel 7006 can be connected to the elution reaction chamber 6010 on the extraction chip 6 through the control valve 2, thereby realizing the transfer of the elution reaction liquid from the extraction chip 6 to the amplification chip 7.

[0046] Please also see Figure 1 and Figure 5The nucleic acid detection kit provided in this embodiment also includes a top cover 1, which is made of a polymer material and is positioned on the extraction chip 6. It can be fixed to the extraction chip 6 by various means, such as double-sided tape, screws, heat pressing, laser or ultrasonic welding, to achieve a sealed top surface of the extraction chip 6. The top cover 1 is provided with injection holes 1001, 1002, 1003, and 1015, and vent holes 1008, 1009, and 1010 for reagent infusion and ventilation. Specifically, the injection hole 1001 is connected to the lysis and combination chamber 6001 for perfusing the lysis solution; the injection hole 1002 is connected to the first cleaning reaction chamber 6002 for perfusing the first cleaning solution; the injection hole 1003 is connected to the second cleaning reaction chamber 6003 for perfusing the second cleaning solution; the injection hole 1015 is connected to the elution reaction chamber 6010 for perfusing the eluent; the ventilation holes 1008, 1009 and 1010 are respectively connected to the lysis and combination chamber 6001, the first cleaning reaction chamber 6002 and the second cleaning reaction chamber 6003 for ventilation. First scale lines 1011, 1012, 1013, and 1014 are provided on the top cover 1. First scale line 1011 is used to indicate the initial position of the rotary valve 4, and first scale line 1014 is used to indicate the initial position of the control valve 2. First scale lines 1011, 1012, and 1013 correspond to the lysis and binding chamber 6001, the first cleaning reaction chamber 6002, and the second cleaning reaction chamber 6003, respectively. This allows precise control of the required rotation angle of the rotary valve 4 during the nucleic acid extraction and purification process, ensuring that the rotary valve 4 and each reaction chamber are aligned and fully sealed, thereby preventing leakage that may affect the test results. In other embodiments, the top cover 1 can be made transparent so that the operator can directly observe the alignment of the rotary valve 4 and the reaction chamber. The first scale lines 1011, 1012, and 1013 may also be omitted from the top cover 1. The top cover 1 is provided with through holes 1004 and 1006 for providing pressure and installation through holes 1005 and 1007. The through hole 1006 is connected with the through hole 6012 for providing pressure on the extraction chip 6. The through hole 1004 is connected with the through hole 6005 for providing pressure on the extraction chip 6 for the piston to pass through. The through hole 1005 is for the installation of the control valve 2. The through hole 1007 is for the installation of the rotary valve 4.

[0047] See Figure 6The control valve 2 includes a second rotating shaft 2001, a third scale line 2002 is set on the side of one end of the second rotating shaft 2001, a microchannel 2003 is set on the end surface of the other end of the second rotating shaft 2001, and a second flange 2004 is formed on the outer side of the second rotating shaft 2001. By rotating the second rotating shaft 2001 to control the direction of the microchannel 2003 on the control valve 2, the elution reaction chamber 6010 is connected with the amplification reaction chambers 7007, 7008, 7009 and 7010. Third scale mark 2002 aligns with first scale mark 1014 on top cover 1, indicating its initial position. At this point, microfluidic channel 2003 disconnects elution reaction chamber 6010 from amplification reaction chambers 7007, 7008, 7009, and 7010. Rotating control valve 2 90° connects microfluidic channel 2003 to elution reaction chamber 6010 and amplification reaction chambers 7007, 7008, 7009, and 7010. Furthermore, the device includes a retaining ring 3 and a second sealing gasket 5 for use with control valve 2. The retaining ring 3 and the second sealing gasket 5 are matingly assembled, with control valve 2 secured within the retaining ring 3. The second sealing gasket 5 is connected to the bottom of the first groove 6007. The second sealing gasket 5 is made of a soft material, such as silicone. A second groove is defined on the second sealing gasket 5, and a protrusion is defined on the retaining ring 3. The protrusion is inserted into the second groove, and a second flange 2004 is matingly connected to the retaining ring 3. A circular hole communicating with the through hole 6009 on the extraction chip 6 is provided on the second sealing gasket 5 . The circular hole is also communicated with the through hole 6008 on the extraction chip 6 and the through hole 7005 on the amplification chip 7 .

[0048] See Figure 7 A first sealing film 8 is set between the extraction chip 6 and the amplification chip 7 to seal the bottom surface of the extraction chip 6 and the top surface of the amplification chip 7. The first sealing film 8 is composed of a polymer and can be combined with the extraction chip 6 and the amplification chip 7 by gluing, hot pressing, laser or ultrasonic welding and maintain a certain strength. A through hole 8001 and air vents 8002, 8003, 8004 and 8005 are provided on the first sealing film 8. The through hole 8001 is communicated with the through hole 6008 on the extraction chip 6 and the through hole 7005 on the amplification chip 7, so that the elution reaction liquid can flow from the elution reaction chamber 6010 of the extraction chip 6 into the amplification reaction chamber 7007-7010 of the amplification chip 7. The air vents 8002, 8003, 8004 and 8005 are communicated with the air vents 7011, 7012, 7013 and 7014 on the amplification chip 7. The area of ​​the first sealing film 8 provided with the air vents 8002, 8003, 8004 and 8005 is located outside the extraction chip 6, that is, the extraction chip 6 does not block this part of the area, so that the air vents 8002, 8003, 8004 and 8005 can be directly ventilated to the outside.

[0049] See Figure 8A second sealing film 9 is disposed at the bottom of the amplification chip 7. This second sealing film 9 is composed of a polymer and can be bonded to the amplification chip 7 by adhesive bonding, heat pressing, laser welding, or ultrasonic welding, maintaining a certain strength. The second sealing film 9 is primarily used to seal the amplification reaction chambers 7007, 7008, 7009, and 7010. Other structures are not particularly limited. Preferably, through-holes 9001, 9002, and 9003 of the same shape as those of the cleavage and binding chamber 6001, the first cleaning reaction chamber 6002, and the second cleaning reaction chamber 6003 are disposed correspondingly.

[0050] See Figure 9 The packaged nucleic acid detection kit mainly includes a top cover 1, an extraction chip 6 and an amplification chip 7.

[0051] Please continue to see Figure 1 、 Figure 2a and Figure 2b The lysis and binding chamber 6001 on the extraction chip 6 encapsulates 600ul of lysis solution containing magnetic beads, the first cleaning reaction chamber 6002 encapsulates the first cleaning solution, the second cleaning reaction chamber 6003 encapsulates the second cleaning solution, and the elution reaction chamber 6010 encapsulates the elution solution.

[0052] Please continue to see Figure 4 Different freeze-dried reaction reagents are encapsulated in the four amplification reaction chambers 7007, 7008, 7009 and 7010 of the amplification detection chip 7 to achieve multiple nucleic acid detection.

[0053] See Figure 10 After chip packaging is complete, the second scale mark 40013 on the rotary valve 4 is aligned with the first scale mark 1011 on the top cover 1, and the third scale mark 2002 on the control valve 2 is aligned with the first scale mark 1014 on the top cover 1. After the sample to be tested is injected into the lysis and binding chamber 6001 of the extraction chip 6 through the injection port 1001, the injection ports 1001, 1002, 1003, 1015 and the vents 1008, 1009, and 1010 on the top cover 1 are sealed with double-sided tape.

[0054] See Figure 2a and Figure 2b, the nucleic acid extraction process of the kit provided in this embodiment is as follows: the kit is loaded into the supporting automated instrument, the automated nucleic acid extraction process is started, the permanent magnetic head on the instrument automatically sticks to the center of the lysis reaction chamber 6001, and absorbs and gathers the magnetic beads; the ultrasonic head ultrasonicates at a certain frequency and amplitude for a certain time to fully lyse the sample to be tested and fully combine with the magnetic beads, and the heating module continuously heats the lysis reaction chamber 6001 at the same time; after the lysis reaction is completed, the permanent magnetic head absorbs the magnetic beads bound to the nucleic acid and transfers them to the extraction port 40021 on the rotary valve 4, and the rotary valve 4 is rotated 60° clockwise, and the magnetic beads are transferred to the first cleaning reaction chamber 6002; the permanent magnetic head automatically sticks to the center of the first cleaning reaction chamber 6002, and absorbs and gathers the magnetic beads; the ultrasonic head ultrasonicates at a certain frequency and amplitude for 1 minute to evenly disperse the magnetic beads to complete the first cleaning reaction; the permanent magnetic head absorbs the magnetic beads bound to the nucleic acid and transfers them to the extraction port 40021 on the rotary valve 4, and the rotary valve is rotated 60° clockwise. 4 is rotated 60° clockwise, and the magnetic beads are transferred to the second cleaning reaction chamber 6003, and the first cleaning step is repeated once; after completing the second cleaning step, the permanent magnetic head adsorbs the magnetic beads bound to the nucleic acid and transfers them to the extraction port 40021 on the rotary valve 4, and the rotary valve 4 is rotated 60° clockwise to transfer them to the first drying chamber 6013. The piston on the instrument can input a certain positive and negative pressure to the first drying chamber 6013 through the through hole 1004, the piston through hole 6005, and the microchannels 6006 and 6016 to generate gas flow, thereby drying the magnetic beads; after drying, the magnetic beads can remain relatively dry and the bound nucleic acid will not be destroyed. The rotary valve 4 is rotated 60° clockwise again, and the magnetic beads are transferred to the elution reaction chamber 6010; the permanent magnetic head adsorbs and aggregates the magnetic beads, and the ultrasonic head sonicates at a certain frequency and amplitude for 4 minutes to evenly disperse the magnetic beads. At the same time, the heating module continuously heats the elution reaction chamber 6010 at 65°C to complete the elution and magnetic bead separation steps.

[0055] See Figure 11 Before the elution step is completed, microchannel 2003 on control valve 2 is closed and disconnected from through-holes 6008 and 7005. At this point, extraction chip 6 and amplification chip 7 are disconnected. After the elution and magnetic bead separation steps are complete, the permanent magnetic head is again positioned against the center of elution reaction chamber 6010 to absorb the immobilized magnetic beads. Control valve 2 is rotated clockwise 90°, connecting microchannel 2003 to through-holes 6008 and 7005. Elution reaction chamber 6010 is now connected to amplification chip 7. A piston applies a positive pressure to elution reaction chamber 6010 from through-holes 1006 and 6012 via microchannels 6011 and 6015, pushing 80 μl of reagents after the elution reaction and magnetic bead separation into amplification chip 7. Control valve 2 is rotated clockwise 90°, sealing microchannel 2003 and isolating extraction chip 6 from amplification chip 7.

[0056] Please continue to see Figure 7 and Figure 11 Different freeze-dried amplification reagents are placed in the four amplification reaction chambers 7007, 7008, 7009 and 7010 on the amplification chip 7, and a waterproof and breathable membrane is provided corresponding to the vent holes 7011, 7012, 7013 and 7014 in each chamber. The vent holes 7011, 7012, 7013 and 7014 are connected to the vent holes 8002, 8003, 8004 and 8005 on the first sealing membrane 8 respectively; after waiting for the elution reaction and the reagents for magnetic bead separation to completely fill the four amplification reaction chambers 7007, 7008, 7009 and 7010 and the microfluidic channel 7006, the vent holes 8002, 8003, 8004 and 8005 are sealed with double-sided tape.

[0057] Finally, PCR amplification and detection are completed under the action of the external instrument thermal cycle module and fluorescence detection module.

[0058] In summary, the nucleic acid detection kit provided by the present invention has the following advantages:

[0059] 1. By packaging the extraction chip, amplification chip, rotary valve, and control valve into one, nucleic acid detection reagents can be integrated into a microfluidic chip; the kit has a compact structure, small size, easy to carry, flexible use, and can adapt to a variety of environments.

[0060] 2. The extraction chip is sealed by the top cover and the first sealing film, and the amplification chip is sealed by the first sealing film and the second sealing film. The entire process of nucleic acid detection, nucleic acid extraction, purification, amplification, detection, etc. can be carried out in a closed space.

[0061] 3. By setting up multiple amplification reaction chambers on the amplification chip and encapsulating different freeze-dried reaction reagents, multiple nucleic acid detection is achieved in one chip system, which improves the nucleic acid detection throughput and reduces the amount of detection samples.

[0062] 4. The nucleic acid detection kit provided by the present invention cooperates with an external control instrument to realize fully automatic nucleic acid detection. All processes of nucleic acid detection are completed automatically in a relatively short time, with fewer operating steps, which reduces personnel time and reduces the errors and uncertainties caused by manual operation to nucleic acid detection, thereby improving the accuracy and stability of nucleic acid detection. At the same time, it can avoid various problems such as reagent contamination and environmental pollution during the nucleic acid detection process.

[0063] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art may make some modifications and improvements without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention shall be based on the definition of the claims.

Claims

1. A nucleic acid detection kit, characterized in that: It includes an extraction chip, a top cover, an amplification chip and a control valve packaged into one body. The extraction chip includes a plurality of chambers arranged in an annular direction and a rotary valve arranged in a rotational direction, wherein some of the chambers are lysis and binding chambers, cleaning reaction chambers, and elution reaction chambers. The rotary valve is provided with an extraction port, which communicates with one of the chambers as the rotary valve rotates. The top cover is arranged on the extraction chip, and a plurality of injection holes, a plurality of ventilation holes and a pressure-providing through hole are arranged on the top cover, the plurality of injection holes are respectively communicated with the cleavage combination chamber, the cleaning reaction chamber and the elution reaction chamber, the plurality of ventilation holes are respectively communicated with the cleavage combination chamber and the cleaning reaction chamber, and the pressure-providing through hole is communicated with the elution reaction chamber through a microchannel; a plurality of first scale lines are arranged on the top cover, each chamber corresponds to a first scale line, a second scale line is arranged corresponding to the rotary valve, and a third scale line is arranged corresponding to the control valve; the rotary valve includes a first rotating shaft and a first sealing gasket, one end of the first rotating shaft is sleeved with the first sealing gasket, the extraction port is arranged on the first sealing gasket, and the other end of the first rotating shaft is arranged with a second scale line; the first rotating shaft has a cavity for accommodating a magnet, a circle of flange is arranged on the outside of the first rotating shaft, the inner side surface of the first sealing gasket is in contact with the first rotating shaft, and the end surface of the first sealing gasket is in contact with the flange; The amplification chip is stacked with the extraction chip, and the amplification chip has an amplification reaction chamber. The control valve is used to control the connection or disconnection of the elution reaction chamber and the amplification reaction chamber, so that the object to be detected is transferred to the amplification reaction chamber; the control valve includes a second rotating shaft, the third scale line is set on the side of one end of the second rotating shaft, and a microchannel is set on the end face of the other end of the second rotating shaft. The direction of the microchannel on the control valve is controlled by rotating the second rotating shaft to connect or disconnect the elution reaction chamber and the amplification reaction chamber.

2. The nucleic acid detection kit according to claim 1, characterized in that The extraction chip is provided with a lysis and combination chamber, a first cleaning reaction chamber, a second cleaning reaction chamber, a first drying chamber, an elution reaction chamber, and a second drying chamber in sequence along the rotation direction of the rotary valve. The second drying chamber isolates the elution reaction chamber from the lysis and combination chamber. The first drying chamber and the second drying chamber are both connected to a through hole for providing pressure.

3. The nucleic acid detection kit according to claim 2, characterized in that The cleavage and combination chamber, the first cleaning reaction chamber, the second cleaning reaction chamber, and the elution reaction chamber are arranged from the top surface to the bottom surface of the extraction chip; the first drying chamber and the second drying chamber are arranged on the back bottom surface of the extraction chip and do not pass through the extraction chip.

4. The nucleic acid detection kit according to claim 1, characterized in that It includes a fixing ring and a second sealing gasket. The extraction chip has a first groove. The control valve is arranged in the first groove. The fixing ring and the second sealing gasket are matched and assembled. The control valve is fixed in the fixing ring. The second sealing gasket is connected to the bottom of the first groove.

5. The nucleic acid detection kit according to claim 1, characterized in that The amplification chip is provided with four amplification reaction chambers for placing different amplification reagents, and the four amplification reaction chambers are connected through a microfluidic channel.

6. The nucleic acid detection kit according to claim 5, characterized in that Each amplification reaction chamber is provided with a vent hole, and a waterproof breathable membrane is provided corresponding to the vent hole.

7. The nucleic acid detection kit according to claim 1, characterized in that It comprises a first sealing film and a second sealing film. The first sealing film is arranged between the extraction chip and the amplification chip, and the bottom surface of the amplification chip is attached to the second sealing film.

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