Nucleic acid extraction device and extraction method

Abstract

The invention discloses a nucleic acid extraction device and method, and relates to the technical field of nucleic acid extraction device.The nucleic acid extraction device comprises a nucleic acid storage table, an isolation hood is fixedly connected to the middle of the upper surface of the nucleic acid storage table, and a preheating dispersion barrel is arranged on the circumference of the upper surface of the nucleic acid storage table; a nucleic acid detection rod is arranged at the upper end of the multifunctional sampling tube; the device further comprises a uniform dispersion preheating adjusting mechanism and an auxiliary sampling mechanism; by arranging the uniform dispersion preheating adjusting mechanism, shaking control over the interior of the specimen can be achieved, by arranging the cambered surface of the bevel convex angle block, the rotating disc can push the preheating dispersion barrel to slide in the isolation hood in a reciprocating mode, and by controlling the rotating speed of the motor, the reciprocating sliding frequency of the preheating dispersion barrel in the isolation hood can be directly controlled; according to the method, sample damage caused by too large physical force generated in the shaking process can be reduced, especially for fragile cells or tissue structures, so that the integrity and extraction efficiency of nucleic acid extraction can be improved.

Description

Technical Field

[0001] The present invention relates to the technical field of nucleic acid extraction devices, in particular to a nucleic acid extraction device and an extraction method. Background Art

[0002] Nucleic acid testing of pigs is an efficient and accurate detection method, which plays an important role in the diagnosis, monitoring and prevention of pig diseases. Nucleic acid testing can be used to diagnose various pig diseases such as swine fever, African swine fever, pseudorabies, etc. By detecting the DNA or RNA of pathogens in pigs, it can be accurately determined whether pigs are infected with related viruses. During the peak period of the epidemic or in the epidemic-spreading area, by regularly collecting pig samples for nucleic acid testing, the epidemic dynamics can be discovered in time, providing important data support for epidemic prevention and control.

[0003] However, when the existing pig nucleic acid is tested, when the sampled nucleic acid test stick is stored, the nucleic acid test stick needs to be placed in a multifunctional sampling tube, the handle needs to be broken, and it needs to be clearly marked. When the operator breaks the handle, he must ensure that the position of the sample tube nucleic acid test stick in the container multifunctional sampling tube is neither offset nor unstable. This is because any slight shaking may affect the smooth breaking of the handle and may even damage the sample tube, resulting in sample contamination or failure to successfully break the handle. However, the existing collection tube design has not fully taken this into consideration. The existing collection tube has a relatively simple design and lacks sufficient stability and support, so that the sample tube nucleic acid test stick is easily shaken during the process of breaking the handle. This shaking not only increases the difficulty of breaking the handle, but also greatly increases the risk of sample contamination.

[0004] Existing pig nucleic acid specimens need to be manually shaken by operators after collection. During the shaking process, the cell walls or tissue structures in the samples will be broken by the physical force, making the nucleic acid in the cells easier to extract. Moderate shaking can also help remove impurities in the sample and improve the purity of the nucleic acid. However, if the physical force generated during the shaking process is too large, it may cause damage to the sample, especially for those fragile cells or tissue structures, which will affect the integrity and extraction efficiency of the nucleic acid. If a separate instrument is used for shaking, additional operating steps and time are required, which may increase the workload of the experimenter. Especially when dealing with a large number of samples, this cumbersome operating process may reduce work efficiency.

[0005] Therefore, a nucleic acid extraction device and an extraction method are proposed to solve the above problems. Summary of the invention

[0006] In view of this, the technical problem to be solved by the present invention is to propose a nucleic acid extraction device and an extraction method to solve the problems of easy contamination during collection of pig nucleic acid samples in the prior art and low detection efficiency caused by single mixing and shaking after sample collection.

[0007] To achieve the above-mentioned object, the present invention provides the following technical solutions: a nucleic acid extraction device and an extraction method, comprising a nucleic acid storage table, an isolation cover is fixedly connected to the middle of the upper surface of the nucleic acid storage table, a preheating dispersion cylinder is arranged on the circumference of the upper surface, a multifunctional sampling tube is arranged in the preheating dispersion cylinder, a nucleic acid detection rod is arranged at the upper end of the multifunctional sampling tube, and also includes a uniform dispersion preheating adjustment mechanism and an auxiliary sampling mechanism, the uniform dispersion preheating adjustment mechanism is arranged in the middle of the isolation cover, and the auxiliary sampling mechanism is arranged in the multifunctional sampling tube;

[0008] The uniform dispersion preheating adjustment mechanism is used for uniform mixing and heating with reagents after nucleic acid extraction;

[0009] The auxiliary sampling mechanism is used to assist in the extraction of nucleic acids.

[0010] Preferably, the uniformly dispersed preheating adjustment mechanism includes a protective cover, the bottom of the protective cover is fixedly connected to the upper surface of the isolation cover, a motor is installed in the middle of the upper surface of the protective cover, and the motor drive shaft passes through the middle of the protective cover, and a rotating drum is rotatably connected to the middle of the upper end of the isolation cover, and the middle of the upper surface of the rotating drum is fixedly connected to the motor drive shaft.

[0011] Preferably, an eccentric hole is provided at an eccentric position of one end of the rotating drum away from the motor, a conical sliding column is slidably connected in the eccentric hole, and a turntable is fixedly connected to one end of the conical sliding column away from the eccentric hole, the turntable is arranged on the inner wall, and an inclined convex corner block is provided on the side of the turntable away from the eccentric hole, and the inclined convex corner block is fixedly connected to the bottom of the isolation cover on one side away from the turntable.

[0012] Preferably, the ends of the preheating dispersion cylinder are evenly arranged on the upper surface of the turntable, a buffer spring is sleeved on the outer surface of the preheating dispersion cylinder near the inner wall of the isolation cover, one end of the buffer spring is fixedly connected to the preheating dispersion cylinder, and the other end of the buffer spring is fixedly connected to the inner wall of the isolation cover, a copper power-on ring is sleeved on the outer surface of the preheating dispersion cylinder near the top of the isolation cover, a shock-absorbing block is slidably connected to the inner wall of the preheating dispersion cylinder, a first reset spring is fixedly connected to the bottom of the shock-absorbing block, and one end of the first reset spring away from the shock-absorbing block is fixedly connected to the bottom of the preheating dispersion cylinder.

[0013] Preferably, an anti-falling mechanism is further provided at the upper end of the preheating dispersion cylinder, and the anti-falling mechanism is used to prevent the multifunctional sampling tube from detaching, and the anti-falling mechanism includes a multifunctional clamping and prying plate, and a support plate is rotatably connected to the middle part of the multifunctional clamping and prying plate, and the support plate is rotatably connected to the outer surface of the upper end of the preheating dispersion cylinder at one end away from the multifunctional clamping and prying plate, and a pressing plate is fixedly connected to one end of the multifunctional clamping and prying plate, and the other end of the multifunctional clamping and prying plate is fixedly connected to a splint, and a slot is provided at the upper end of the preheating dispersion cylinder, and a second reset spring is evenly and fixedly connected in the slot, and the second reset spring is fixedly connected to the splint of the multifunctional clamping and prying plate at one end away from the slot.

[0014] Preferably, the auxiliary sampling mechanism includes a retaining ring, which is fixedly connected to the inner wall of the upper end of the multifunctional sampling tube, and a limiting hole is provided on the retaining ring. A sealing cover is rotatably connected to the outer surface of the upper end of the multifunctional sampling tube, and a non-slip limiting plate is fixedly connected to the side of the sealing cover away from the connection with the multifunctional sampling tube.

[0015] Preferably, the nucleic acid detection stick is arranged below the retaining ring near the nucleic acid extraction end, an auxiliary groove is provided on the nucleic acid detection stick, and the auxiliary groove of the nucleic acid detection stick is arranged at the upper end of the multifunctional sampling tube.

[0016] The nucleic acid extraction device and the extraction method include the following steps:

[0017] Step 1: Sampling of clinical specimens: Use the nucleic acid detection stick to scrape the inside cheeks of the pig's mouth and the nasal cavity. If necessary, such as anal swabs or vaginal swabs, they must also be collected according to the corresponding methods. Then put the nucleic acid detection stick into the multi-functional sampling tube, break off the handle, and mark it clearly;

[0018] Step 2: Extraction of nucleic acid: Extract viral RNA or DNA from the sample. RNA can be reverse transcribed (convert RNA into cDNA) to obtain a certain number of DNA or cDNA molecules.

[0019] Step 3: Multiplex quantitative PCR detection and nanopore sequencing library construction; after 1.5 hours of multiplex quantitative PCR detection, a graphical curve display can be obtained on the terminal. After the nanopore sequencing library is constructed after 10 minutes, the ends of these DNA or cDNA molecules are repaired and connected to the sequencing adapter to complete the library construction, and then placed in the sequencing chip. The process of molecules entering and passing through the nanopore is monitored in real time through the electrical signal feedback system and the sequence information of each molecule is recorded;

[0020] Step 4, Minion sequencing: basic preprocessing of the data obtained through sequencing, such as filtering low-quality data, removing the adapter sequences after sequencing, splicing and assembling the DNA or cDNA sequences, etc.; finally, the data results are compared with the known gene library, and the virus infection situation is identified (Centrifuge software analysis) and abnormal genes are discovered (R language analysis).

[0021] Compared with the prior art, the present invention provides a nucleic acid extraction device and extraction method, which have the following beneficial effects:

[0022] 1. Through the setting of the retaining ring and the anti-slip limiting plate in the present invention, by placing the nucleic acid detection end of the nucleic acid detection stick at the bottom of the retaining ring and in the limiting hole, the limiting hole and the anti-slip limiting plate limit the detection rod of the nucleic acid detection stick, so that the detection rod can be stably tilted at the upper end of the multi-functional sampling tube. At this time, when the sealing cover is closed, the auxiliary groove of the nucleic acid detection stick is pressed and engaged by the anti-slip limiting plate. At this time, the operator can easily break the handle of the nucleic acid detection stick. Compared with the prior art, through the setting of the present invention, the operator only needs to align the nucleic acid detection stick with the limiting hole and then press the sealing cover to easily break the handle. This can not only avoid the specimen falling and contamination phenomenon caused by the sliding of the nucleic acid detection stick when breaking in the prior art, but also reduce the operator's control over the breaking of the nucleic acid detection stick, enhance the operator's stability and anti-falling when breaking the handle of the nucleic acid detection stick, and thus reduce the pollution phenomenon during specimen collection.

[0023] 2. The uniform dispersion preheating adjustment mechanism can not only control the internal shaking of the specimen, but also enable the turntable to push the preheating dispersion cylinder to slide back and forth in the isolation cover through the arc surface setting of the inclined convex corner block. By controlling the rotation speed of the motor, the frequency of the reciprocating sliding of the preheating dispersion cylinder in the isolation cover can be directly controlled, which can not only reduce the excessive physical force generated during the shaking process, resulting in damage to the sample, especially for those fragile cells or tissue structures, but also increase the integrity and efficiency of nucleic acid extraction.

[0024] Moreover, by placing the multifunctional sampling tube in a preheated dispersion cylinder for storage, not only can the contamination from external pollution be reduced, but the rotation of the turntable can also drive multiple collected samples to be properly shaken, further ensuring that the cells or tissues in the sample can be effectively lysed, thereby releasing nucleic acids. In addition, by stably clamping the surface of the multifunctional sampling tube through the multifunctional clamping pry plate, the multifunctional sampling tube can be stably stored in the preheated dispersion cylinder during transportation, thereby reducing the occurrence of problems such as falling and loss of collected samples.

[0025] 3. According to the invention, a copper energized ring is sleeved on the outer surface of the preheating dispersion tube. When the preheating dispersion tube slides back and forth on the isolation cover, in order to denature the protein in the sample, thereby destroying the structure of the cell wall and the cell membrane, the nucleic acid in the cell is more easily released, and a moderate heating method is used for nucleic acid extraction. By passing current into multiple copper energized rings connected in series, more accurate and efficient heating is achieved, and a temperature sensor is installed near the multifunctional sampling tube, which can monitor and control the internal temperature in real time, thereby ensuring that the heating process is both fast and stable. Compared with the traditional heating method, the present invention greatly shortens the heating time, and the temperature control is more accurate, reducing the risk of nucleic acid degradation caused by temperature fluctuations. More importantly, through the design of the present invention, we can not only achieve batch, controllable temperature heating treatment, but also shake the specimen during the heating process, and this shaking is conducive to uniform heating inside the sample, so that the nucleic acid can be released more quickly, thereby further improving the efficiency of nucleic acid extraction. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is a schematic diagram of the three-dimensional structure of the present invention;

[0027] Figure 2 It is a half-cut auxiliary schematic diagram of the three-dimensional structure of the present invention;

[0028] Figure 3 It is a half-cut schematic diagram of the connection relationship of the partial structure of the uniform dispersion preheating adjustment mechanism of the present invention;

[0029] Figure 4 It is a schematic diagram of the structural connection relationship of the anti-falling mechanism of the present invention;

[0030] Figure 5 For the present invention Figure 4 Enlarged view of point A in the middle;

[0031] Figure 6 This is a schematic diagram of the structural connection relationship of the auxiliary sampling mechanism of the present invention;

[0032] Figure 7 For the present invention Figure 6 Enlarged view of point B in the middle.

[0033] In the figure:

[0034] 1. Nucleic acid storage table; 11. Isolation cover; 12. Preheating dispersion cylinder; 13. Multifunctional sampling tube; 14. Nucleic acid detection stick;

[0035] 2. Uniformly dispersed preheating adjustment mechanism; 21. Protective cover; 22. Rotating drum; 23. Eccentric hole; 24. Rotating plate; 25. Inclined convex corner block; 26. Buffer spring; 27. Shock absorber block; 28. First reset spring; 29. ​​Copper power ring;

[0036] 3. Anti-fall mechanism; 31. Multifunctional clamping and prying plate; 32. Support plate; 33. Second return spring;

[0037] 4. Auxiliary sampling mechanism; 41. Retaining ring; 42. Limiting hole; 43. Sealing cover; 44. Anti-slip limiting plate. DETAILED DESCRIPTION

[0038] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0039] The present invention is further described in detail below based on the accompanying drawings and embodiments.

[0040] First embodiment

[0041] Please refer to Figures 1 to 7 As shown:

[0042] In order to solve the problems mentioned in the technical solution, the embodiment of the present application provides a nucleic acid extraction device and an extraction method, including a nucleic acid storage table 1, an isolation cover 11 is fixedly connected to the middle of the upper surface of the nucleic acid storage table 1, a preheating dispersion cylinder 12 is arranged on the circumference of the upper surface, a multifunctional sampling tube 13 is arranged in the preheating dispersion cylinder 12, a nucleic acid detection rod 14 is arranged at the upper end of the multifunctional sampling tube 13, and also includes a uniform dispersion preheating adjustment mechanism 2 and an auxiliary sampling mechanism 4, the uniform dispersion preheating adjustment mechanism 2 is arranged in the middle of the isolation cover 11, and the auxiliary sampling mechanism 4 is arranged in the multifunctional sampling tube 13;

[0043] The uniform dispersion preheating adjustment mechanism 2 is used for uniform mixing and heating of the reagents after nucleic acid extraction;

[0044] The auxiliary sampling mechanism 4 is used to assist in the extraction of nucleic acids;

[0045] The uniform dispersion preheating adjustment mechanism 2 includes a protective cover 21, the bottom of the protective cover 21 is fixedly connected to the upper surface of the isolation cover 11, a motor is installed in the middle of the upper surface of the protective cover 21, and the motor drive shaft passes through the middle of the protective cover 21, and a rotating drum 22 is rotatably connected to the middle of the upper end of the isolation cover 11, and the middle of the upper surface of the rotating drum 22 is fixedly connected to the motor drive shaft;

[0046] An eccentric hole 23 is provided at an eccentric position of one end of the rotating drum 22 away from the motor, a conical sliding column is slidably connected in the eccentric hole 23, and a rotating disk 24 is fixedly connected to one end of the conical sliding column away from the eccentric hole 23, the rotating disk 24 is arranged on the inner wall, and an inclined convex corner block 25 is arranged on one side of the rotating disk 24 away from the eccentric hole 23, and the inclined convex corner block 25 is fixedly connected to the bottom of the isolation cover 11 on one side away from the rotating disk 24;

[0047] The ends of the preheating dispersion cylinder 12 are evenly arranged on the upper surface of the rotating disk 24, and a buffer spring 26 is sleeved on the outer surface of the preheating dispersion cylinder 12 near the inner wall of the isolation cover 11. One end of the buffer spring 26 is fixedly connected to the preheating dispersion cylinder 12, and the other end of the buffer spring 26 is fixedly connected to the inner wall of the isolation cover 11. A copper power ring 29 is sleeved on the outer surface of the preheating dispersion cylinder 12 near the top of the isolation cover 11, and a shock-absorbing block 27 is slidably connected to the inner wall of the preheating dispersion cylinder 12. A first reset spring 28 is fixedly connected to the bottom of the shock-absorbing block 27, and one end of the first reset spring 28 away from the shock-absorbing block 27 is fixedly connected to the bottom of the preheating dispersion cylinder 12;

[0048] Among them, the copper power rings 29 arranged on the circumference of the upper surface of the isolation cover 11 are connected in series and fixed to the non-metallic baffle, and a temperature sensor device is arranged on the circumference of the baffle to detect the internal temperature of the preheating dispersion cylinder 12.

[0049] Compared with the prior art, through the implementation of this embodiment, the present invention greatly shortens the heating time, and the temperature control is more precise, reducing the risk of nucleic acid degradation caused by temperature fluctuations. More importantly, through the design of the present invention, we can not only achieve batch, controllable temperature heating treatment, but also shake the specimen during the heating process. This shaking is conducive to uniform heating inside the sample, allowing the nucleic acid to be released more quickly, thereby further improving the efficiency of nucleic acid extraction.

[0050] For further examples, please refer to Figures 1 to 7 As shown:

[0051] The upper end of the preheating dispersion tube 12 is also provided with an anti-falling mechanism 3, which is used to prevent the multifunctional sampling tube 13 from falling off, and the anti-falling mechanism 3 includes a multifunctional clamping pry plate 31, the middle part of the multifunctional clamping pry plate 31 is rotatably connected with a support plate 32, and the support plate 32 is rotatably connected to the outer surface of the upper end of the preheating dispersion tube 12 at one end away from the multifunctional clamping pry plate 31, and a pressing plate is fixedly connected to one end of the multifunctional clamping pry plate 31, and the other end of the multifunctional clamping pry plate 31 is fixedly connected to a clamping plate, and a card slot is opened at the upper end of the preheating dispersion tube 12, and a second reset spring 33 is evenly fixedly connected in the card slot, and the second reset spring 33 is fixedly connected to the clamping plate of the multifunctional clamping pry plate 31 at one end away from the card slot;

[0052] The auxiliary sampling mechanism 4 includes a retaining ring 41, which is fixedly connected to the inner wall of the upper end of the multifunctional sampling tube 13. A limiting hole 42 is provided on the retaining ring 41. A sealing cover 43 is rotatably connected to the outer surface of the upper end of the multifunctional sampling tube 13. A non-slip limiting plate 44 is fixedly connected to the side of the sealing cover 43 away from the connection with the multifunctional sampling tube 13.

[0053] The nucleic acid detection rod 14 is arranged below the retaining ring 41 near the nucleic acid extraction end, and an auxiliary groove is provided on the nucleic acid detection rod 14. The auxiliary groove of the nucleic acid detection rod 14 is arranged at the upper end of the multifunctional sampling tube 13;

[0054] Among them: the rotating connection between the sealing cover 43 and the multifunctional sampling tube 13 is adapted to the size of the card slot opened at the upper end of the preheating dispersion tube 12, and the limiting blocks are fixedly connected on both sides of the bottom of the retaining ring 41 to prevent the nucleic acid detection plate 14 from sliding when the nucleic acid is extracted.

[0055] Compared with the prior art, through the implementation of this embodiment, the operator only needs to tilt and align the nucleic acid detection stick 14 with the limiting hole 42, and then press the sealing cover 43 to easily break the handle. This can not only avoid the sample falling and contamination phenomenon caused by the sliding of the nucleic acid detection stick 14 when breaking in the prior art, but also reduce the operator's control over the breaking of the nucleic acid detection stick 14, enhance the operator's stability and anti-falling when breaking the handle of the nucleic acid detection stick 14, and thus reduce the contamination phenomenon during sample collection.

[0056] Second embodiment

[0057] The nucleic acid extraction device and the extraction method comprise the following steps:

[0058] Step 1, sampling of clinical specimens: Use the nucleic acid detection stick 14 to scrape the inside cheeks of the pig's mouth and scrape the nasal cavity. If necessary, such as anal swabs or vaginal swabs, they must also be collected according to the corresponding methods, and then the nucleic acid detection stick 14 is placed in the multifunctional sampling tube 13, the handle is broken, and it is clearly marked;

[0059] Step 2, extraction of nucleic acid: extract viral RNA or DNA from the sample, reverse transcribe the RNA to convert it into cDNA, and obtain a certain number of DNA or cDNA molecules;

[0060] Step 3: Multiplex quantitative PCR detection and nanopore sequencing library construction; after 1.5 hours of multiplex quantitative PCR detection, a graphical curve display can be obtained on the terminal. After the nanopore sequencing library is constructed after 10 minutes, the ends of these DNA or cDNA molecules are repaired and connected to the sequencing adapter to complete the library construction, and then placed in the sequencing chip. The process of molecules entering and passing through the nanopore is monitored in real time through the electrical signal feedback system and the sequence information of each molecule is recorded;

[0061] Step 4, Minion sequencing: basic preprocessing of the data obtained through sequencing, such as filtering low-quality data, removing the sequence of the adapter after sequencing, splicing and assembling the DNA or cDNA sequence, etc.; finally, the data results are compared with the known gene library, and the virus infection status and abnormal gene analysis are identified through Centrifuge software analysis and R language analysis;

[0062] Compared with traditional sequencing technology, the nanopore sequencing technology in this method has the following characteristics:

[0063] ① Longer sequencing read length: Nanopore sequencing technology uses the change of electrical signals when bases pass through nanopores to achieve sequencing. In principle, it can detect all nucleic acid sequences passing through the nanopore. Therefore, there is no limit on the sequencing length. Currently, the longest read length can exceed 2M;

[0064] ② Real-time sequencing: The sequencing library preparation process is simple, and there is no need to perform PCR amplification on the nucleic acid in the sample. DNA or RNA can be sequenced directly, saving operation time and reducing sequencing costs. The results can be output while sequencing, enabling real-time analysis of sequencing data;

[0065] ③ Direct RNA sequencing: Nanopore sequencing can directly sequence all forms of RNA, which can avoid the bias and possible mutations caused by reverse transcription of RNA into DNA amplification when sequencing and studying RNA viruses. This feature is of great appeal in the field of RNA virus detection;

[0066] Everything in the above example works like this:

[0067] In the initial state: the motor is not started, the multifunctional sampling tube 13 is not placed in the preheating dispersion tube 12, and the copper power ring 29 is not energized.

[0068] The following is the working process of the auxiliary sampling mechanism 4 for assisting the extraction of nucleic acid:

[0069] When collecting nucleic acid specimens from imported pigs, a special nucleic acid detection stick 14 and a multifunctional sampling tube 13 are used. The sampling personnel need to wear protective clothing and gloves to ensure aseptic operation. At this time, the operator uses the collection end of the nucleic acid detection stick 14 to scrape the inside cheeks of the pig's mouth and the inside of the nasal cavity. If necessary, such as anal swabs or vaginal swabs, they must also be collected according to the corresponding methods. Then, the collection end of the nucleic acid detection stick 14 is placed in the inner wall of the multifunctional sampling tube 13 below the retaining ring 41 and the handle is broken. Figure 7As shown, by placing the nucleic acid detection end of the nucleic acid detection stick 14 at the bottom of the retaining ring 41 and in the limiting hole 42, the limiting hole 42 and the anti-slip limiting plate 44 limit the detection rod of the nucleic acid detection stick 14, so that the detection rod can be stably tilted at the upper end of the multi-functional sampling tube 13. At this time, when the sealing cover 43 is closed, the anti-slip limiting plate 44 presses and engages the auxiliary groove of the nucleic acid detection stick 14. At this time, the operator can easily break the handle of the nucleic acid detection stick 14. At this time, the broken handles are collected uniformly, and the specimen collection end located in the multi-functional sampling tube 13 is passed through the nucleic acid detection stick 14. After breaking, the specimen begins to fall into the bottom of the multifunctional sampling tube 13 to be mixed with the reagent. Compared with the prior art, the present invention only requires the operator to tilt and align the nucleic acid detection stick 14 with the limiting hole 42, and then press the sealing cover 43 to easily break the handle. This can not only avoid the specimen falling and contamination caused by the sliding of the nucleic acid detection stick 14 when breaking in the prior art, but also reduce the operator's control over the breaking of the nucleic acid detection stick 14, enhance the operator's stability and anti-falling when breaking the handle of the nucleic acid detection stick 14, and thus reduce the contamination during specimen collection.

[0070] Please refer to the above working process Figures 1 to 7 .

[0071] The following is the working process of the uniform dispersion preheating adjustment mechanism 2 for uniform mixing and heating with reagents after nucleic acid extraction:

[0072] As described above, after the specimens are uniformly collected, the operator uniformly puts the multifunctional sampling tube 13 into the preheated dispersion tube 12, and then gently presses the multifunctional sampling tube 13, such as Figure 4 Figure 5As shown, at this time, by pressing the nucleic acid detection stick 14, the operator places the multifunctional sampling tube 13 and the slot opened in the preheating dispersion tube 12 on the same motion trajectory. When the multifunctional sampling tube 13 slides into the slot opened in the preheating dispersion tube 12, since one end of the multifunctional clamping pry plate 31 is fixedly connected to the pressing plate, the other end of the multifunctional clamping pry plate 31 is fixedly connected to the clamping plate, and the second return spring 33 is evenly fixedly connected in the slot, and the second return spring 33 is fixedly connected to the clamping plate of the multifunctional clamping pry plate 31 away from one end of the slot, when the connection of the sealing cover 43 at the upper end of the multifunctional sampling tube 13 is gradually squeezed to the clamping plate, the squeezing of the clamping plate will drive the second return spring 33 to be compressed, and at this time the bottom of the multifunctional sampling tube 13 In contact with the upper surface of the shock-absorbing block 27, the multifunctional sampling tube 13 can be stably stored in the preheating dispersion cylinder 12 through the elastic action of the second return spring 33 in the clamp; by placing the multifunctional sampling tube 13 in the preheating dispersion cylinder 12 for storage, not only can the contamination from external pollution be reduced, but the rotation of the turntable 24 can also drive multiple collected samples to be properly shaken, further ensuring that the cells or tissues in the samples can be effectively lysed, thereby releasing the nucleic acid, and by stably clamping the surface of the multifunctional sampling tube 13 with the multifunctional clamping pry plate 31, the multifunctional sampling tube 13 can be stably stored in the preheating dispersion cylinder 12 during the transportation process, thereby reducing the occurrence of problems such as the falling and loss of the collected samples.

[0073] At this time, in order to speed up the decomposition of nucleic acids, the operator can start the switch button and temperature control switch for controlling the start of the motor on the nucleic acid storage table 1, and set the optimal nucleic acid extraction temperature according to different specimens. At this time, the motor installed on the upper surface of the protective cover 21 starts to rotate under the electrical control of the switch. Because the motor drive shaft passes through the protective cover 21 and is fixedly connected to the middle part of the upper end of the drum 22, the drum 22 will start to rotate in the same direction when the motor rotates. An eccentric hole 23 is provided at the eccentric part of the bottom of the drum 22, and a conical sliding column fixedly connected to the middle part of the turntable 24 is slidably connected in the eccentric hole 23. Moreover, the inclined convex corner block 25 is fixedly connected to the bottom of the isolation cover 11, and the middle part of the turntable 24 is arranged on the inclined convex corner block 25. Since the inclined convex corner block 25 adopts a shoe upper conical setting, At this time, the rotation of the rotating cylinder 22 will drive the conical sliding column to drive the turntable 24 to rotate with different slopes on the inclined convex corner block 25. Furthermore, because the preheating dispersion cylinder 12 slides in a circle in the isolation cover 11, and the bottom of the preheating dispersion cylinder 12 is arranged on the upper surface of the turntable 24, the preheating dispersion cylinder 12 will slide in the isolation cover 11 through the rotation of the turntable 24 with different amplitudes. At this time, the multifunctional sampling tube 13 engaged in the preheating dispersion cylinder 12 begins to shake the specimen inside the multifunctional sampling tube 13 under the movement of the preheating dispersion cylinder 12. By shaking the inside of the multifunctional sampling tube 13 by the preheating dispersion cylinder 12, not only can the damage of the sample caused by excessive physical force generated during the shaking process be reduced, especially for those fragile cells or tissue structures, but also the integrity and efficiency of nucleic acid extraction can be increased.

[0074] At this time, the start-up of the stable control switch and the threshold control of the temperature sensor device enable the copper power ring 29 to quickly heat the preheating dispersion tube 12. The temperature sensor can be used to control the internal temperature of the preheating dispersion tube 12, and then the heat transfer inside the preheating dispersion tube 12 can gradually heat the inside of the multi-functional sampling tube 13. At this time, the preheating dispersion tube 12 slides in the isolation cover 11, and the copper power ring 29 heats it. At the same time, the preheating dispersion tube 12 slides back and forth in the copper power ring 29, so that the inside of the preheating dispersion tube 12 can be evenly heated. The copper power ring 29 is sleeved on the outer surface of the preheating dispersion tube 12. When the preheating dispersion tube 12 slides back and forth on the isolation cover 11, in order to denature the protein in the sample, the structure of the cell wall and cell membrane is destroyed, making the nucleic acid in the cell more easily released. Release it, so as to adopt a moderate heating method to extract nucleic acid. By passing current into multiple copper power rings 29 connected in series, more accurate and efficient heating can be achieved, and a temperature sensor is installed near the multifunctional sampling tube 13, which can monitor and control the internal temperature in real time, so as to ensure that the heating process is both fast and stable. Compared with the traditional heating method, the design of the present invention not only greatly shortens the heating time, but also makes the temperature control more accurate, reducing the risk of nucleic acid degradation caused by temperature fluctuations. More importantly, through the design of the present invention, we can not only realize batch and controllable temperature heating treatment, but also make the specimen shake during the heating process, and this shaking is conducive to uniform heating inside the sample, so that the nucleic acid can be released more quickly, thereby further improving the efficiency of nucleic acid extraction.

[0075] Please refer to the above working process Figures 1 to 7 .

[0076] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0077] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A nucleic acid extraction device for nucleic acid collection and access, comprising a nucleic acid storage table (1), wherein an isolation cover (11) is fixedly connected to the middle of the upper surface of the nucleic acid storage table (1), a preheating dispersion cylinder (12) is arranged on the circumference of the upper surface, a multifunctional sampling tube (13) is arranged in the preheating dispersion cylinder (12), and a nucleic acid detection rod (14) is arranged at the upper end of the multifunctional sampling tube (13), characterized in that: It also includes a uniform dispersion preheating adjustment mechanism (2) and an auxiliary sampling mechanism (4), wherein the uniform dispersion preheating adjustment mechanism (2) is arranged in the middle of the isolation cover (11), and the auxiliary sampling mechanism (4) is arranged in the multifunctional sampling tube (13); The uniform dispersion preheating adjustment mechanism (2) is used for uniform mixing and heating of nucleic acid and reagents after nucleic acid extraction; The auxiliary sampling mechanism (4) is used to assist in the extraction of nucleic acids.

2. A nucleic acid extraction device according to claim 1, characterized in that: The uniform dispersion preheating adjustment mechanism (2) comprises a protective cover (21), the bottom of the protective cover (21) is fixedly connected to the upper surface of the isolation cover (11), a motor is installed in the middle of the upper surface of the protective cover (21), and the motor drive shaft passes through the middle of the protective cover (21), a rotating drum (22) is rotatably connected to the middle of the upper end of the isolation cover (11), and the middle of the upper surface of the rotating drum (22) is fixedly connected to the motor drive shaft.

3. A nucleic acid extraction device according to claim 2, characterized in that: An eccentric hole (23) is provided at an eccentric position of one end of the rotating drum (22) away from the motor, a conical sliding column is slidably connected in the eccentric hole (23), and a rotating disk (24) is fixedly connected to one end of the conical sliding column away from the eccentric hole (23), the rotating disk (24) is arranged on the inner wall, and an inclined convex corner block (25) is provided on one side of the rotating disk (24) away from the eccentric hole (23), and the inclined convex corner block (25) is fixedly connected to the bottom of the isolation cover (11) on one side away from the rotating disk (24).

4. A nucleic acid extraction device according to claim 3, characterized in that: The ends of the preheating dispersion cylinder (12) are evenly arranged on the upper surface of the rotating disk (24); a buffer spring (26) is sleeved on the outer surface of the preheating dispersion cylinder (12) near the inner wall of the isolation cover (11); one end of the buffer spring (26) is fixedly connected to the preheating dispersion cylinder (12); the other end of the buffer spring (26) is fixedly connected to the inner wall of the isolation cover (11); a copper power ring (29) is sleeved on the outer surface of the preheating dispersion cylinder (12) near the top of the isolation cover (11); a shock absorbing block (27) is slidably connected to the inner wall of the preheating dispersion cylinder (12); a first return spring (28) is fixedly connected to the bottom of the shock absorbing block (27); and one end of the first return spring (28) away from the shock absorbing block (27) is fixedly connected to the bottom of the preheating dispersion cylinder (12).

5. A nucleic acid extraction device according to claim 1, characterized in that: The upper end of the preheating dispersion tube (12) is also provided with an anti-falling mechanism (3), which is used to prevent the multifunctional sampling tube (13) from falling off, and the anti-falling mechanism (3) includes a multifunctional clamping and prying plate (31), the middle part of which is rotatably connected with a support plate (32), and the support plate (32) is rotatably connected to the outer surface of the upper end of the preheating dispersion tube (12) at one end away from the multifunctional clamping and prying plate (31), and a pressing plate is fixedly connected to one end of the multifunctional clamping and prying plate (31), and the other end of the multifunctional clamping and prying plate (31) is fixedly connected to a clamping plate, and a slot is provided at the upper end of the preheating dispersion tube (12), and a second reset spring (33) is evenly and fixedly connected in the slot, and the second reset spring (33) is fixedly connected to the clamping plate of the multifunctional clamping and prying plate (31) at one end away from the slot.

6. A nucleic acid extraction device according to claim 1, characterized in that: The auxiliary sampling mechanism (4) comprises a retaining ring (41), the retaining ring (41) is fixedly connected to the inner wall of the upper end of the multifunctional sampling tube (13), a limiting hole (42) is provided on the retaining ring (41), a sealing cover (43) is rotatably connected to the outer surface of the upper end of the multifunctional sampling tube (13), and a non-slip limiting plate (44) is fixedly connected to the side of the sealing cover (43) away from the connection with the multifunctional sampling tube (13).

7. A nucleic acid extraction device according to claim 1, characterized in that: The nucleic acid detection stick (14) is arranged below the retaining ring (41) near the nucleic acid extraction end, and an auxiliary groove is provided on the nucleic acid detection stick (14). The auxiliary groove of the nucleic acid detection stick (14) is arranged at the upper end of the multifunctional sampling tube (13).

8. A method for extracting nucleic acid using a nucleic acid extraction device, applicable to a nucleic acid extraction device according to any one of claims 1 to 7, characterized in that: The following steps are involved: Step 1, sampling of clinical specimens: Use a nucleic acid detection stick (14) to scrape the inside cheeks of the pig's mouth and scrape the inside of the nasal cavity. If necessary, such as anal swabs or vaginal swabs, they must also be collected according to the corresponding methods. Then, the nucleic acid detection stick (14) is placed in a multifunctional sampling tube (13), the handle is broken, and it is clearly marked; Step 2: Extraction of nucleic acid: Extract viral RNA or DNA from the sample. RNA can be reverse transcribed (convert RNA into cDNA) to obtain a certain number of DNA or cDNA molecules. Step 3: Multiplex quantitative PCR detection and nanopore sequencing library construction: After 1.5 hours of multiplex quantitative PCR detection, a graphical curve display can be obtained on the terminal. After 10 minutes of nanopore sequencing library construction, the ends of these DNA or cDNA molecules are repaired and connected to sequencing adapters to complete the library construction, and then placed in the sequencing chip. The electrical signal feedback system monitors the process of molecules entering and passing through the nanopore in real time and records the sequence information of each molecule; Step 4, MinION sequencing: basic preprocessing of the data obtained through sequencing. In step 3, low-quality data is filtered out, the adapter sequences after sequencing are removed, and the DNA or cDNA sequences are spliced ​​and assembled. Finally, the data results are compared with the known gene library, and the virus infection status is identified (Centrifuge software analysis) and abnormal genes are discovered (R language analysis).

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