Microorganism detection and identification device and identification method

Abstract

The invention relates to the field of microbiological detection, in particular to a microbiological detection and identification device and identification method.The microbiological detection and identification device comprises a main cylinder, a bearing plate is slidably arranged in the middle of the main cylinder, a plurality of circular grooves are formed in the bearing plate, storage units are placed in the circular grooves, and a puncture assembly is arranged in a lower cavity; one side of the puncture assembly extends into the circular groove and the storage unit and is used for supplementing an extraction reagent into the circular groove and the storage unit, a limiting assembly used for limiting and plugging the storage unit is arranged at the upper end of the main cylinder, and a nanopore tester connected with the puncture assembly is mounted and inserted in the bottom of the main cylinder; through mutual cooperation of the storage unit, the puncture assembly and the adsorption assembly, when a microbial sample is detected, microbial DNA and RNA nucleic acid separation can be efficiently achieved, nucleic acid is secondarily extracted through the adsorption assembly, the purity of the nucleic acid sample is accurately improved, and the detection efficiency is improved. The problems of low extraction purity and more impurities in the traditional method are solved.

Description

Technical Field

[0001] The present invention relates to the field of microbial detection, and in particular to a microbial detection and identification device and an identification method. Background Art

[0002] Microbial detection and identification devices are automated devices used to rapidly identify microbial species in samples. By integrating core steps such as nucleic acid extraction, separation, purification, and sequencing analysis, they automate the entire process from sample processing to result output. This technology can be applied in clinical diagnosis, food hygiene monitoring, and environmental microbiology analysis, effectively improving detection efficiency and reducing the risk of contamination caused by manual operations, providing an efficient and standardized solution for microbial testing.

[0003] The current microbial detection technology system has obvious technological generational limitations: in terms of operational procedures, it relies on a serial operation mode of discrete equipment such as centrifuges and nucleic acid extractors, which requires samples to undergo multiple manual transfers and interface exposures; in terms of contamination control, the open operating architecture exposes samples repeatedly to the laboratory environment, posing a significant risk of biological contamination; in terms of automation, existing technologies are unable to achieve an end-to-end closed loop from sample input to result output, and key links such as lysate transfer and magnetic bead purification still rely heavily on manual operations, restricting detection efficiency and result consistency.

[0004] Prior art, such as patent publication number CN109813695A, discloses a microfluidic chip-based microbial detection system and method. These methods involve enabling the flow of samples and reagents through the microchannels of the microfluidic chip, and utilizing a standing wave acoustic field (including acoustic radiation force, lateral acoustic force, and interactive acoustic force) generated by an ultrasonic device to gather microorganisms in an observation area for optical detection. However, while this technology has improved some of the original issues, there are still areas that require further optimization to better meet actual detection needs.

[0005] 1. This technology does not have an automated continuous detection process, which means that it cannot continuously detect different test objects in a cycle, cannot meet the needs of real-time online monitoring in industrial production, and the equipment has low reusability.

[0006] 2. Its microchannel adopts a single flat capillary structure, which has large fluid resistance and easily causes uneven distribution of microorganisms during the flow process, affecting the ultrasonic aggregation effect. At the same time, it lacks anti-clogging design, and long-term use can easily lead to microchannel blockage due to particle deposition.

[0007] Therefore, based on the above-stated viewpoint, there is still room for optimization in the existing technology for the extraction and detection of microorganisms. Summary of the Invention

[0008] In order to solve the above problems, the present invention provides a microorganism detection and identification device, including a main cylinder, a support plate is slidably arranged in the middle of the main cylinder, and the support plate divides the main cylinder into an upper chamber and a lower chamber, a plurality of circular grooves are opened on the support plate, a storage unit for storing extracts is placed in the circular grooves, and a puncture assembly is arranged in the lower chamber, one side of the puncture assembly extends into the circular groove and the storage unit and is used to replenish the extraction reagent therein.

[0009] An adsorption component is also provided in the circular groove for adsorbing the DNA and RNA of microorganisms in the extraction reagent.

[0010] A limiting component for limiting and sealing the storage unit is provided at the upper end of the main cylinder.

[0011] A vibration component is arranged on the supporting plate.

[0012] A nanopore tester connected to the puncture component is installed at the bottom of the main tube and is used to extract the detection solution in the circular groove through the puncture component.

[0013] Preferably, the storage unit includes a tank body placed in a circular groove, an inner tank is slidably inserted inside the tank body, and a plurality of strip grooves distributed along its axis are opened on the outside of the inner tank, an upper cover plate is provided on the inner tank by a threaded connection, and the outer side of the upper cover plate extends to the upper end of the tank body, and at the same time the through-opening of the inner tank and the tank body is sealed, and a structural groove is also opened on the inner wall of the tank body.

[0014] Preferably, a through groove is provided at the bottom of the tank body, and a blocking block extending into the through groove is provided at the bottom of the inner tank.

[0015] Preferably, the puncture assembly includes a plurality of puncture grooves provided on the support plate and corresponding to the through grooves one by one, and a plurality of puncture shafts corresponding to the puncture grooves are provided on the bottom wall of the lower chamber. The top end of the puncture shaft passes through the puncture groove and extends into the corresponding circular groove and contacts the bottom of the blocking block. A conveying groove is provided in the puncture shaft, one side of which extends to the top thereof, and a bending groove is provided in the blocking block, the bottom of which corresponds to and passes through the conveying groove, and the other end of the bending groove passes through the outside of the blocking block and is connected to the inside of the tank body. A pipetting groove is provided on the outside of the puncture shaft and the outside of the blocking block.

[0016] Preferably, the puncture shaft is provided with an infusion groove with both ends extending to the outside thereof, the outside of the puncture shaft is connected to a bending tube, and one side of the bending tube extends to the nanopore tester and is connected to its test port.

[0017] Several liquid storage boxes corresponding to the puncture shafts are also installed in the lower chamber. One side of the liquid storage box is penetrated by an infusion tube with one end penetrating into the inside of the puncture shaft and communicating with the delivery groove.

[0018] A push plate is slidably provided in the liquid storage box, and push screw rods which are rotatably penetrated on both sides of the liquid storage box and are threadedly connected with the push plate.

[0019] Preferably, the adsorption component includes two clips symmetrically arranged in the circular groove, and the test paper is clamped between the clips.

[0020] Preferably, the limiting assembly includes a sealing sleeve installed at the upper end of the main cylinder, and a plurality of rings corresponding to the circular grooves are provided on the inner side of the sealing sleeve. An upper plate sliding in the ring contacts the corresponding upper cover plate, and the upper plate and the sealing sleeve are connected by a reset push spring.

[0021] Preferably, an annular ring is provided on the inner bottom wall of the circular groove, and the end face of the annular ring is in contact with the corresponding tank body. One end of the annular ring is provided with several limiting shafts that slide through the bottom of the tank body. An annular groove is also provided on the inner bottom wall of the circular groove, and an active push spring is provided between the inner bottom wall of the annular groove and the annular ring.

[0022] Preferably, the vibration assembly includes a support plate arranged on the upper end of the support plate, a high-frequency vibration motor is installed on the support plate, an extension frame is provided on the outside of the vibration motor, and the ends of several extension sections of the extension frame are installed with transfer rings mounted on the outside of the corresponding tank body.

[0023] In addition, the present invention also provides a method for detecting and identifying microorganisms, comprising the following steps: S1, sample processing: The collected sample is placed in a storage unit, which is then placed in a circular groove. The extraction reagent is then injected into the sample through the puncture component to separate the microbial DNA and RNA nucleic acids to obtain a mixed solution.

[0024] S2, nucleic acid extraction: the mixed solution then enters the circular groove, the adsorption component in the circular groove adsorbs and extracts the nucleic acid, and the remaining mixed solution is discharged from the circular groove by the puncture component.

[0025] S3, nucleic acid solution transfer: New extraction reagent is injected into the circular groove through the puncture component again to mix the nucleic acid attached to the adsorption component to obtain a nucleic acid solution with higher purity. The nucleic acid solution is transported to the nanopore tester through the puncture component.

[0026] S4, sequencing by nanopore sequencer: Use the nanopore tester to sequence the nucleic acid solution. The nanopore tester starts the process of sequencing and analyzing at the same time. After obtaining enough data to determine the high-quality sequencing data, a test report is issued according to the actual situation. Sequencing is continued according to actual needs, and test reports are issued under different sequencing data amounts for verification and comparison.

[0027] In summary, this application includes at least one of the following beneficial technical effects: 1. The present invention cooperates with the storage unit, puncture component and adsorption component to efficiently separate microbial DNA and RNA nucleic acids during microbial sample testing, and uses the adsorption component to extract nucleic acids for the second time, accurately improving the purity of nucleic acid samples and solving the problems of low extraction purity and high impurities in traditional methods. 2. The present invention utilizes the characteristics of the nanopore tester that performs sequencing and analysis simultaneously, combined with clear data quality and positive judgment criteria, to flexibly and quickly issue a test report after obtaining sufficiently high-quality sequencing data, without having to complete the entire sequencing process, greatly shortening the test cycle and improving detection efficiency. 3. The present invention realizes the automation of operations such as support plate movement and extraction reagent delivery through the coordinated operation of various transmission components in the drive unit, reduces manual intervention and reduces human errors. At the same time, the drive unit cooperates with other components to further improve the degree of automation and accuracy of the entire microbial detection and identification process. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The present invention will be further described below with reference to the accompanying drawings and examples.

[0029] Figure 1 It is a structural diagram of the main body of the present invention.

[0030] Figure 2 It is a cross-sectional view of the main body of the present invention.

[0031] Figure 3 It is a cross-sectional view of the storage unit of the present invention.

[0032] Figure 4 This invention Figure 3 A magnified view of part of the structure at point A.

[0033] Figure 5 It is a cross-sectional view of the puncture assembly of the present invention.

[0034] Figure 6 It is a structural schematic diagram of the adsorption component of the present invention.

[0035] Figure 7 This invention Figure 6 A magnified view of part of the structure at point B.

[0036] Figure 8 It is a cross-sectional view of the limiting component and the vibration component of the present invention.

[0037] Figure 9 It is a structural schematic diagram of the drive unit of the present invention.

[0038] Figure 10 It is a diagram showing the coordination of the arc-shaped magnetic plate and the passive magnetic plate of the present invention.

[0039] Figure 11It is a diagram showing the coordination of the vertical shaft, horizontal shaft, drive shaft and reciprocating screw rod of the present invention.

[0040] In the figure, 1, main cylinder; 10, supporting plate; 11, circular groove; 12, nanopore tester; 2, storage unit; 20, tank body; 21, inner tank; 22, strip groove; 23, upper cover; 24, structural groove; 25, blocking block; 3, puncture assembly; 30, puncture groove; 31, puncture shaft; 32, conveying groove; 33, bending groove; 34, pipetting groove; 35, infusion groove; 36, bending tube; 37, liquid storage box; 38, infusion tube; 39, push plate; 310, Push screw; 4. Adsorption assembly; 40. Clip; 41. Test paper; 5. Limit assembly; 50. Sealing sleeve; 51. Ring; 52. Upper plate; 53. Annular ring; 54. Limit shaft; 6. Vibration assembly; 60. Vibration motor; 61. Transfer ring; 7. Drive unit; 70. Drive cavity; 71. Vertical slot; 72. Vertical axis; 73. Horizontal axis; 74. Drive shaft; 75. Arc slot; 76. Reciprocating screw; 77. Arc magnetic plate; 78. Passive magnetic plate. DETAILED DESCRIPTION

[0041] The following combination Figures 1 to 11 The embodiments of the present invention are described in detail.

[0042] The embodiments of the present application disclose a microbial detection and identification device and identification method. The present application is applied in the process of detecting and identifying microbial samples. Through innovative structural design, it can efficiently separate microbial DNA and RNA nucleic acids and accurately extract nucleic acid samples, solving the problems of cumbersome procedures and low extraction purity of traditional methods. Furthermore, the present application can also use a nanopore tester to achieve sequencing and analysis at the same time, flexibly issue a test report based on strict data quality and positive judgment standards, and use a drive unit to realize automatic operation of the device, significantly improving the efficiency and accuracy of detection and identification, and providing a new technical solution for the field of microbial detection.

[0043] Example 1: Reference Figure 1 、 Figure 2 and Figure 3 As shown, it includes a main cylinder 1, a supporting plate 10, a circular groove 11, a storage unit 2, a puncture component 3, an adsorption component 4, a limit component 5, a vibration component 6 and a nanopore tester 12. A supporting plate 10 is slidably arranged in the middle of the main cylinder 1, and the supporting plate 10 divides the main cylinder 1 into an upper chamber and a lower chamber. A plurality of circular grooves 11 are opened on the supporting plate 10, and a storage unit 2 for storing an extract is placed in the circular groove 11, and a puncture component 3 is arranged in the lower chamber, and one side of the puncture component 3 extends into the circular groove 11 and the storage unit 2 and is used to replenish the extraction reagent therein.

[0044] An adsorption component 4 is also provided in the circular groove 11 for adsorbing DNA and RNA of microorganisms in the extraction reagent.

[0045] A limiting assembly 5 for limiting and sealing the storage unit 2 is provided at the upper end of the main cylinder 1 .

[0046] A vibration assembly 6 is provided on the supporting plate 10 .

[0047] A nanopore tester 12 connected to the puncture assembly 3 is installed at the bottom of the main tube 1 and is used to extract the detection solution in the circular groove 11 through the puncture assembly 3.

[0048] In addition, the present invention also provides a method for detecting and identifying microorganisms, comprising the following steps: S1, sample processing: the collected sample is placed in the storage unit 2, and then the storage unit 2 is placed in the circular groove 11, and then the extraction reagent is injected into the sample through the puncture component 3, so that the microbial DNA and RNA nucleic acids are separated to obtain a mixed solution.

[0049] S2, extracting nucleic acid: the mixed solution then enters the circular groove 11, and the adsorption component 4 in the circular groove 11 adsorbs and extracts the nucleic acid, and the remaining mixed solution is discharged from the circular groove 11 by the puncture component 3.

[0050] S3, nucleic acid solution transfer: inject new extraction reagent into the circular groove 11 through the puncture component 3 again to mix the nucleic acid adsorbed on the adsorption component 4 to obtain a nucleic acid solution with higher purity. The nucleic acid solution is transported to the nanopore tester 12 through the puncture component 3.

[0051] S4, nanopore sequencer sequencing: Use the nanopore tester 12 to sequence the nucleic acid solution. Nanopore sequencing starts the process of sequencing and analyzing at the same time. After obtaining sufficient data to determine high-quality sequencing data, a test report is issued according to the actual situation, and sequencing is continued according to actual needs. Test reports are issued under different sequencing data amounts for verification and comparison.

[0052] Reference Figure 3 and Figure 4 As shown, a storage unit 2 for storing the extract is placed therein; specifically, the storage unit 2 includes a tank body 20, an inner tank 21, a strip groove 22, an upper cover plate 23, a structural groove 24 and a blocking block 25. The tank body 20 is placed inside the circular groove 11, and the inner tank 21 is slidably inserted inside the tank body 20, and a plurality of strip grooves 22 distributed along the axis thereof are provided on the outer side of the inner tank 21. An upper cover plate 23 is provided on the inner tank 21 by a threaded connection, and the outer side of the upper cover plate 23 extends to the upper end of the tank body 20, while blocking the through-openings of the inner tank 21 and the tank body 20.

[0053] A structural groove 24 is also provided on the inner side wall of the tank body 20 .

[0054] A through groove is formed at the bottom of the tank body 20 , and a blocking block 25 extending into the through groove is provided at the bottom of the inner tank 21 .

[0055] During actual use, a collection material such as a swab or test paper for collecting microorganisms (for the convenience of subsequent explanation, swabs or test paper are collectively referred to as collection materials) is placed in the inner tank 21, and then the upper cover plate 23 is threadedly inserted into the top of the inner tank 21, and then the inner tank 21 is installed in the tank body 20, and the tank body 20 is placed in the circular groove 11. After the inner tank 21 is inserted into the tank body 20, the blocking block 25 will simultaneously block the through groove. At this time, an extraction reagent is injected into the tank body 20, and the extraction reagent dilutes the collection material. After the reaction occurs, the microbial nucleic acid in the collection material will enter the extraction reagent.

[0056] Reference Figure 3 As shown, the puncture assembly 3 is used to supplement the extraction reagent inside the storage unit 2; specifically, the puncture assembly 3 includes a puncture groove 30, a puncture shaft 31, a conveying groove 32, a bending groove 33, a pipette groove 34, an infusion groove 35, a bending tube 36, a liquid storage box 37, an infusion tube 38, a pushing plate 39 and a pushing screw 310. Several puncture grooves 30 are opened on the supporting plate 10 and correspond to the through grooves one by one. Several puncture shafts 31 corresponding to the puncture grooves 30 are provided on the bottom wall of the lower chamber. The top of the puncture shaft 31 passes through the puncture groove 3 0 extends into the corresponding circular groove 11 and contacts the bottom of the blocking block 25, and in actual operation, a sealing ring (known technology) can be installed on the puncture shaft 31 and the bottom of the blocking block 25 to prevent liquid leakage, and it is a routine operation for those skilled in the art, so it will not be repeated here; a conveying groove 32 is provided in the puncture shaft 31, one side of which extends to the top thereof, and a bending groove 33 is provided in the blocking block 25, the bottom of which corresponds to and passes through the conveying groove 32, and the other end of the bending groove 33 passes through the outside of the blocking block 25 and is connected to the interior of the tank body 20.

[0057] That is, when the support plate 10 drives the tank body 20 to descend in the main cylinder 1, the support plate 10 is movable compared to the puncture shaft 31, so the puncture shaft 31 can enter the circular groove 11 through the puncture groove 30, and then contact the bottom of the blocking block 25, and the bottom of the bending groove 33 will be connected with the top of the conveying groove 32.

[0058] A pipetting groove 34 is provided on the outside of the puncture shaft 31 and the blocking block 25 .

[0059] The puncture shaft 31 is provided with an infusion groove 35 with both ends extending to the outside thereof. A bending tube 36 is connected to the outside of the puncture shaft 31 , and one side of the bending tube 36 extends to the nanopore tester 12 and is connected to its test port.

[0060] Several liquid storage boxes 37 corresponding to the puncture shafts 31 are also installed in the lower chamber. One side of the liquid storage box 37 is penetrated by an infusion tube 38 with one end penetrating into the interior of the puncture shaft 31 and communicating with the delivery groove 32.

[0061] A push plate 39 is slidably provided in the liquid storage box 37 , and push screws 310 are rotatably provided on both sides of the liquid storage box 37 and are threadedly connected to the push plate 39 .

[0062] During actual use, the pushing screw 310 is driven by external force to drive the pushing plate 39 to move in the liquid storage box 37. The liquid storage box 37 is used to store the extraction reagent. The pushing plate 39 can drive the extraction reagent from the infusion tube 38 into the conveying groove 32. The extraction reagent then enters the bending groove 33 through the conveying groove 32 and is finally discharged into the tank body 20. Thereafter, the extraction reagent enters the inner tank 21 from the structural groove 24 and the strip groove 22 to contact and mix with the extract, so that the nucleic acid in the extract is extracted by the extraction reagent. At this time, the nucleic acid will spread into the extraction reagent. This is the initial extraction.

[0063] Furthermore, a plurality of external connecting tubes are inserted at the lower end of the main cylinder 1 and are connected to the liquid storage box 37 . The operator can supply the extraction reagent into the liquid storage box 37 through the external connecting tubes.

[0064] After the initial extraction is completed, the support plate 10 continues to descend, so that the puncture shaft 31 continues to drive the inner tank 21 to separate from the tank body 20 through the blocking block 25, so that the liquid in the inner tank 21 can flow into the tank body 20 through the strip groove 22 and the structural groove 24, while the extract in the inner tank 21 is blocked by the strip groove 22 and cannot move out of the inner tank 21. Then the top of the puncture shaft 31 will gradually move into the tank body 20, and at this time the pipetting groove 34 will correspond to the inside of the tank body 20 and the circular groove 11, that is, the space inside the tank body 20 is connected with the circular groove 11 through the structural groove 24, and the extraction reagent in the tank body 20 will enter the circular groove 11 through the structural groove 24.

[0065] Since the extraction reagent at this time contains not only the nucleic acid of the microorganism but also some impurities of the extract, the adsorption component 4 can perform secondary extraction of the nucleic acid in the extraction reagent. After the extraction is completed, the support plate 10 continues to move downward, so that the upper end of the pipette groove 34 outside the puncture shaft 31 is connected with the circular groove 11, and the extraction reagent in the circular groove 11 will enter the pipette groove 34. Then, the extraction reagent will enter the nanopore tester 12 through the pipette groove 34 and the bent tube 36. At this time, the nanopore tester 12 will not test the extraction reagent but will directly discharge it out of the nanopore tester 12.

[0066] Then the support plate 10 moves upward so that the top of the puncture shaft 31 is located in the circular groove 11, and at this time the liquid storage box 37 again discharges the extraction reagent into the circular groove 11 through the conveying groove 32, so that the extraction reagent extracts the nucleic acid in the adsorption component 4. The nucleic acid extracted this time is pure. After that, the above steps are repeated to discharge the extraction reagent into the nanopore tester 12, and the nucleic acid in the extraction reagent is tested by the nanopore tester 12.

[0067] The nanopore tester 12 can analyze the extracted reagents while sequencing, and can issue analysis results without completing the entire sequencing process; when sufficient data has been obtained to determine high-quality sequencing data, a test report can be issued based on the actual situation, or sequencing can continue according to actual needs, and test reports can be issued at different sequencing data volumes for verification and comparison.

[0068] The data quality judgment criteria are as follows: (1) Ctr11 (external reference sequence 1), the target is active and the number of reads is normal, such as reads <10; (2) Ctr12 (external reference sequence 2), 8 targets are active and the ratio is normal.

[0069] (3) Cleanreads account for >70%.

[0070] (4) The number of aligned cleanreads is >20,000.

[0071] The criteria for judging a positive test result are as follows: (1) The number of amplicons is greater than 3.

[0072] (2) Read count performance, consistent with the standard curve designed for the product (color judgment).

[0073] (3) There is no cross-contamination or aerosol contamination signal interference between samples.

[0074] After the device completes the test, the cleaning operation can be performed according to the following process: remove the tank body 20 from the circular groove 11, inject cleaning liquid into the liquid storage box 37 through the external connecting tube, push the screw 310 to drive the push plate 39, so that the cleaning liquid enters the delivery groove 32 of the puncture shaft 31 through the infusion tube 38, and then flows into the interior of the tank body 20 through the bending groove 33 of the blocking block 25, flushing the nucleic acid impurities remaining on the inner wall of the tank body 20, the structural groove 24 and the surface of the inner tank 21.

[0075] The waste liquid after flushing enters the infusion groove 35 of the puncture shaft 31 through the structural groove 24 and the pipetting groove 34 at the bottom of the circular groove 11, flows into the nanopore tester 12 through the bending tube 36, and is finally discharged from the tester discharge port.

[0076] At the same time, the vibration component 6 vibrates to enhance the flushing effect of the cleaning liquid on the inner wall of the circular groove and the residual stains in the puncture groove 30, ensuring that there is no risk of sample cross-contamination inside the device and preparing for the next test.

[0077] Reference Figure 6 and Figure 7 As shown, the adsorption component 4 is used to adsorb the DNA of microorganisms in the extraction reagent; specifically, the adsorption component 4 includes a clamp 40 and a test paper 41, and the two clamps 40 are symmetrically arranged inside the circular groove 11, and the test paper 41 is clamped between the clamps 40.

[0078] The clips 40 at both ends can clamp the test paper 41. The extraction reagent that enters the circular groove 11 for the first time contains a large amount of nucleic acid. Therefore, the test paper 41 at this time can adsorb part of the nucleic acid in the extraction reagent. When the extraction reagent is discharged from the circular groove 11, there is no nucleic acid in the extraction reagent that enters the circular groove 11 for the second time. Since the test paper 41 has already reacted once, the extraction reagent that enters the circular groove 11 for the second time contacts the test paper 41, and the nucleic acid in the test paper 41 will enter the extraction reagent.

[0079] During actual use, the corresponding test paper 41 is first clamped on the corresponding clip 40 , and then the storage unit 2 is installed in the circular groove 11 .

[0080] Reference Figure 4 and Figure 8 As shown, a limiting assembly 5 for limiting and sealing the storage unit 2 is provided at the upper end of the main tube 1; specifically, the limiting assembly 5 includes a sealing sleeve 50, a collar 51, an upper plate 52, an annular ring 53 and a limiting shaft 54. The sealing sleeve 50 is installed at the upper end of the main tube 1. A plurality of collars 51 corresponding to the circular grooves 11 are provided on the inner side of the sealing sleeve 50. An upper plate 52 in contact with the corresponding upper cover plate 23 slides inside the collar 51. The upper plate 52 and the sealing sleeve 50 are connected by a reset push spring.

[0081] That is, after the tank body 20 is installed in the circular groove 11, the sealing sleeve 50 is installed on the upper end of the main tube 1, and then the upper plate 52 will contact the top of the upper cover plate 23 on the inner tank 21 under the push of the corresponding reset push spring, indirectly limiting the inner tank 21, and when the inner tank 21 is driven by the puncture shaft 31 to move up and down in the tank body 20, the upper plate 52 will also move back and forth on the ring 51 and always resist the inner tank 21 through the upper cover plate 23, so that when the rear puncture shaft 31 no longer resists the inner tank 21, the inner tank 21 can slide into the tank body 20 again.

[0082] An annular ring 53 is provided on the inner bottom wall of the circular groove 11, and the end face of the annular ring 53 is in contact with the corresponding tank body 20. One end of the annular ring 53 is provided with a plurality of limit shafts 54 that are slidably penetrated through the bottom of the tank body 20. An annular groove is also provided on the inner bottom wall of the circular groove 11, and an active push spring is provided between the inner bottom wall of the annular groove and the annular ring 53.

[0083] That is, after the tank body 20 is installed in the circular groove 11, the bottom will contact the annular ring 53. At this time, the annular ring 53 can push the tank body 20 to the upper end of the circular groove 11 under the drive of the corresponding active push spring. The annular ring 53 can leave a certain gap between the tank body 20 and the bottom wall of the circular groove 11 to prevent the bottom of the tank body 20 from colliding with the adsorption component 4, and the limiting shaft 54 ​​can limit the tank body 20 to prevent the tank body 20 from accidentally displacing or rotating on the annular ring 53.

[0084] Continue to refer to Figure 8 As shown, the vibration component 6 is used to further mix the microorganisms in the extraction reagent; specifically, the vibration component 6 includes a vibration motor 60 and a transfer ring 61, the support plate is arranged at the upper end of the support plate 10, and the high-frequency vibration motor 60 is installed on the support plate. An extension frame is provided on the outside of the vibration motor 60, and the ends of several extension sections of the extension frame are equipped with transfer rings 61 that are sleeved on the outside of the corresponding tank body 20, and the transfer ring 61 is slidably connected to the tank body 20.

[0085] When the vibration motor 60 is running, it generates vibrations. These vibrations are transmitted to the tank body 20 through the extension frame and transfer ring 61, promoting a more thorough mixing and reaction between the extraction reagent and the extract. As the inner tank 21 moves upward and the extraction reagent inside flows into the tank body 20 through the structural groove 24, the vibrations accelerate the flow of the solution. Furthermore, during the subsequent adsorption reaction between the extraction reagent and the test strip 41, and during the extraction of nucleic acids from the test strip 41 by the extraction reagent entering the circular groove 11 a second time, the vibrations can also significantly improve the reaction efficiency of each step.

[0086] When the tank body 20 vibrates, the corresponding active push springs on the upper plate 52 and the annular ring 53 can play a buffering and efficiency-enhancing effect, and the limiting shaft 54 ​​at this time can play a limiting effect on the tank body 20.

[0087] Further explanation: The test paper 41 is based on a glass fiber membrane with a surface modified with a silicon-based adsorption material. The silanol groups on its surface can physically adsorb nucleic acids with the help of electrostatic effects. When the mixed solution containing nucleic acids first enters the designated circular groove or other structure, the vibration component 6 is activated, driving the operation of related components, prompting the test paper 41 to fully contact the solution. Relying on the electrostatic adsorption ability of the silanol groups, the nucleic acids in the solution are adsorbed and retained on the test paper 41. Impurities that may exist in the solution are not yet adsorbed by the test paper 41 and remain in the initial mixed solution.

[0088] After the first mixed solution is discharged, an extraction reagent with the same composition but without nucleic acid is injected. At this time, the vibration component 6 continues to work, and the extraction reagent cooperates with the force generated by the vibration to gradually break the electrostatic bond between the silanol group and the nucleic acid. Since the physical adsorption of the test paper 41 is a reversible process and it does not have special chemical modifications or specific binding groups, in this adsorption and desorption cycle, the nucleic acid adsorbed on the test paper 41 will be partially released into the injected extraction reagent solution.

[0089] The key role of this process is to allow the nucleic acid released from the test paper 41 into the solution to be free from the interference of impurities in the first mixed solution as much as possible; although it cannot be guaranteed that the solution contains only nucleic acids, such secondary treatment can greatly reduce the impurity content and improve the accuracy of subsequent tests.

[0090] Example 2: Reference Figure 9 、 Figure 10 and Figure 11 As shown, on the basis of embodiment 1, in order to be able to drive the supporting plate 10 and the supporting plate 10 to move, a driving cavity 70 is opened inside the main cylinder 1, and a driving unit 7 is arranged in the driving cavity 70; specifically, the driving unit 7 includes a driving cavity 70, a vertical slot 71, a vertical shaft 72, a horizontal shaft 73, a driving shaft 74, an arc groove 75, a reciprocating screw rod 76, an arc magnetic plate 77 and a passive magnetic plate 78, and several vertical slots 71 corresponding to the liquid storage box 37 are opened in the main cylinder 1, and the bottom of the vertical slot 71 is connected to the driving cavity 70, and a vertical shaft 72 is rotated inside the vertical slot 71, and several one-to-one magnetic plates are rotatably penetrated on the inner wall of the main cylinder 1. The horizontal shaft 73 is located in the corresponding vertical groove 71, and the horizontal shaft 73 is located on one side of the vertical groove 71 and is connected to the vertical shaft 72 through a gear transmission. The other side of the horizontal shaft 73 passes through the outer wall of the corresponding liquid storage box 37 and is connected to the pushing screw 310. The vertical shaft 72 can rotate in the vertical groove 71, and the vertical shaft 72 can also drive the horizontal shaft 73 to rotate through a gear transmission during the rotation process. When the horizontal shaft 73 rotates, it can drive the corresponding pushing screw 310 to rotate synchronously, so that the pushing screw 310 drives the corresponding pushing plate 39 to move back and forth in the liquid storage box 37.

[0091] A drive shaft 74 is also provided on the inner bottom wall of the drive cavity 70. The bottoms of several vertical shafts 72 extend into the drive cavity 70 and are connected by belt transmission. The vertical shaft 72 on one side is connected to the drive shaft 74 by belt transmission, that is, the vertical shafts 72 can rotate synchronously by gear transmission. When the drive shaft 74 is driven by an external drive device (existing motor equipment), it can drive the vertical shaft 72 on one side to rotate by belt transmission, so that all the vertical shafts 72 are driven to rotate.

[0092] The main cylinder 1 is also provided with an arc-shaped groove 75 whose bottom is connected to the drive cavity 70. A reciprocating screw rod 76 rotates on the top wall of the arc-shaped groove 75, and the outer side of the reciprocating screw rod 76 is threadedly connected to an arc-shaped magnetic plate 77 slidably set in the arc-shaped groove 75, and a passive magnetic plate 78 corresponding to the arc-shaped magnetic plate 77 is also provided on one side of the support plate 10. The bottom of the reciprocating screw rod 76 extends into the drive cavity 70 and is connected to the drive shaft 74 by a belt drive. During the rotation process, the drive shaft 74 can also drive the reciprocating screw rod 76 to rotate by a belt drive. When the reciprocating screw rod 76 rotates, it can also drive the arc-shaped magnetic plate 77 to move up and down in the arc-shaped groove 75. When moving, the arc-shaped magnetic plate 77 can also drive the passive magnetic plate 78 and the support plate 10 connected thereto to move up and down in the main cylinder 1 by a magnetic connection.

[0093] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the present invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the present invention. Therefore, the embodiments should be considered in all respects as exemplary and non-restrictive.

[0094] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.

Claims

1. A microbial detection and identification device, comprising a main cylinder (1), characterized in that: A supporting plate (10) is slidably provided in the middle of the main cylinder (1), and the supporting plate (10) divides the main cylinder (1) into an upper chamber and a lower chamber. A plurality of circular grooves (11) are provided on the supporting plate (10), and a storage unit (2) for storing an extract is placed in the circular groove (11), and a puncture component (3) is provided in the lower chamber. One side of the puncture component (3) extends into the circular groove (11) and the storage unit (2) and is used to replenish the extraction reagent therein. An adsorption component (4) is also provided in the circular groove (11) for adsorbing DNA and RNA of microorganisms in the extraction reagent; A limiting component (5) for limiting and sealing the storage unit (2) is provided at the upper end of the main cylinder (1); A vibration assembly (6) is provided on the supporting plate (10); A nanopore tester (12) connected to the puncture assembly (3) is installed and inserted at the bottom of the main cylinder (1) and is used to extract the detection solution in the circular groove (11) through the puncture assembly (3).

2. A microorganism detection and identification device according to claim 1, characterized in that: The storage unit (2) includes a tank body (20) placed in a circular groove (11), an inner tank (21) is slidably inserted into the tank body (20), and a plurality of strip grooves (22) distributed along the axis of the inner tank (21) are provided on the outer side of the inner tank (21), an upper cover plate (23) is provided on the inner tank (21) by means of a threaded connection, and the outer side of the upper cover plate (23) extends to the upper end of the tank body (20), and at the same time blocks the through-opening of the inner tank (21) and the tank body (20), and a structural groove (24) is also provided on the inner side wall of the tank body (20).

3. A microorganism detection and identification device according to claim 2, characterized in that: A through groove is provided at the bottom of the tank body (20), and a blocking block (25) extending into the through groove is provided at the bottom of the inner tank (21).

4. A microorganism detection and identification device according to claim 3, characterized in that: The puncture assembly (3) includes a plurality of puncture grooves (30) provided on the support plate (10) and corresponding to the through grooves. A plurality of puncture shafts (31) corresponding to the puncture grooves (30) are provided on the bottom wall of the lower chamber. The top end of the puncture shaft (31) passes through the puncture groove (30) and extends into the corresponding circular groove (11) and contacts the bottom of the blocking block (25). A conveying groove (32) extending from one side to the top of the puncture shaft (31) is provided, and a bending groove (33) corresponding to and passing through the conveying groove (32) is provided in the blocking block (25). The other end of the bending groove (33) passes through the outside of the blocking block (25) and is connected to the inside of the tank body (20). A transfer groove (34) is provided on the outside of the puncture shaft (31) and the blocking block (25).

5. A microorganism detection and identification device according to claim 4, characterized in that: The puncture shaft (31) is provided with an infusion groove (35) with both ends extending to the outside thereof. The outside of the puncture shaft (31) is connected to a bending tube (36), and one side of the bending tube (36) extends to the nanopore tester (12) and is connected to its test port. A plurality of liquid storage boxes (37) corresponding to the puncture shafts (31) are also installed in the lower chamber. One side of the liquid storage box (37) is penetrated by an infusion tube (38) having one end extending through the interior of the puncture shaft (31) and connected to the delivery groove (32); A push plate (39) is slidably provided in the liquid storage box (37), and push screw rods (310) are rotatably provided on both sides of the liquid storage box (37) and are threadedly connected to the push plate (39).

6. A microorganism detection and identification device according to claim 1, characterized in that: The adsorption assembly (4) comprises two clips (40) symmetrically arranged in the circular groove (11), and a test paper (41) is clamped between the clips (40).

7. A microorganism detection and identification device according to claim 1, characterized in that: The limiting assembly (5) includes a blocking sleeve (50) mounted on the upper end of the main cylinder (1), a plurality of collars (51) corresponding to the circular grooves (11) are provided on the inner side of the blocking sleeve (50), an upper plate (52) in contact with the corresponding upper cover plate (23) slides inside the collar (51), and the upper plate (52) and the blocking sleeve (50) are connected via a reset spring.

8. A microorganism detection and identification device according to claim 7, characterized in that: An annular ring (53) is provided on the inner bottom wall of the circular groove (11), and the end surface of the annular ring (53) contacts and abuts against the corresponding tank body (20). One end of the annular ring (53) is provided with a plurality of limit shafts (54) that are slidably penetrated through the bottom of the tank body (20). An annular groove is also provided on the inner bottom wall of the circular groove (11), and an active push spring is provided between the inner bottom wall of the annular groove and the annular ring (53).

9. A microorganism detection and identification device according to claim 2, characterized in that: The vibration assembly (6) includes a support plate arranged at the upper end of the support plate (10), a high-frequency vibration motor (60) is mounted on the support plate, an extension frame is arranged outside the vibration motor (60), and transfer rings (61) are mounted on the ends of several extension sections of the extension frame and are sleeved on the outside of the corresponding tank body (20).

10. A microbial detection and identification method, using a microbial detection and identification device according to any one of claims 1 to 9, characterized in that: The identification method includes the following steps: S1, sample processing: placing the collected sample in the storage unit (2), then placing the storage unit (2) in the circular groove (11), and then injecting the extraction reagent into the sample through the puncture component (3), so that the microbial DNA and RNA nucleic acids are separated to obtain a mixed solution; S2, extracting nucleic acid: the mixed solution then enters the circular groove (11), the adsorption component (4) in the circular groove (11) adsorbs and extracts the nucleic acid, and the remaining mixed solution is discharged from the circular groove (11) by the puncture component (3); S3, nucleic acid solution transfer: inject new extraction reagent into the circular groove (11) through the puncture component (3) again to mix the nucleic acid attached to the adsorption component (4) to obtain a nucleic acid solution with higher purity. The nucleic acid solution is transported to the nanopore tester (12) through the puncture component (3); S4, sequencing by nanopore sequencer: The nucleic acid solution is sequenced using a nanopore tester (12). The nanopore tester (12) starts a sequencing and analysis process. After obtaining sufficient data to determine high-quality sequencing data, a test report is issued based on the actual situation. Sequencing is continued based on actual needs, and test reports are issued under different sequencing data volumes for verification and comparison.

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