A microbial detection and identification device and identification method

Abstract

This invention relates to the field of microbial detection, and particularly to a microbial detection and identification device and method. The device includes a main cylinder with a support plate slidably disposed in the middle. Several circular grooves are formed on the support plate, and storage units are placed within these grooves. A puncture assembly is disposed in the lower chamber, with one side extending into the circular grooves and storage units for replenishing extraction reagents. A limiting assembly for limiting and sealing the storage units is disposed at the upper end of the main cylinder. A nanopore analyzer connected to the puncture assembly is installed at the bottom of the main cylinder for extracting the detection solution from the circular grooves through the puncture assembly. This invention, through the cooperation of the storage unit, puncture assembly, and adsorption assembly, can efficiently separate microbial DNA and RNA nucleic acids during microbial sample detection, and utilizes the adsorption assembly for secondary extraction of nucleic acids, accurately improving the purity of nucleic acid samples and solving the problems of low purity and numerous impurities in traditional methods.

Description

Technical Field

[0001] This invention relates to the field of microbial detection, and in particular to a microbial detection and identification device and method. Background Technology

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

[0003] Current microbial detection technologies suffer from significant generational limitations: in terms of operational procedures, the reliance on serial operation modes using discrete equipment such as centrifuges and nucleic acid extractors results in samples undergoing multiple manual transfers and interface exposures; in terms of contamination control, the open operational architecture exposes samples repeatedly to the laboratory environment, posing a significant risk of biocontamination; and in terms of automation, existing technologies cannot achieve an end-to-end closed loop from sample input to result output, with key steps such as lysis buffer transfer and magnetic bead purification still heavily reliant on manual operation, thus limiting detection efficiency and result consistency.

[0004] In the prior art, such as the patent with publication number CN109813695A, a microbial detection system and its detection method based on microfluidic chips are disclosed. This includes realizing the flow of samples and reagents through the microchannels of the microfluidic chip, and using the standing wave sound field (including sound wave radiation force, transverse sound force and interactive sound force) generated by an ultrasonic device to gather microorganisms to the observation area for optical detection. However, although this technology has improved some of the original problems, there are still aspects that need further optimization in order to better meet the actual detection needs.

[0005] 1. This technology is not designed with an automated continuous detection process, meaning it cannot continuously detect different test objects in a loop, thus failing to 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 high fluid resistance and is prone to 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 is prone to microchannel blockage due to particle deposition.

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

[0008] To address the aforementioned problems, the present invention provides a microbial detection and identification device, comprising a main cylinder, a support plate slidably disposed in the middle of the main cylinder, and the support plate dividing the main cylinder into an upper chamber and a lower chamber. The support plate has several circular grooves, in which storage units for storing extracts are placed, and a puncture component is disposed in the lower chamber, one side of which extends into the circular grooves and the storage units and is used to replenish the extraction reagents inside.

[0009] Furthermore, the circular groove is equipped with an adsorption component to adsorb the DNA and RNA of microorganisms from the extraction reagent.

[0010] The upper end of the main cylinder is equipped with a limiting component for limiting and sealing the storage unit.

[0011] Vibration components are installed on the support plate.

[0012] A nanopore tester connected to the puncture assembly is installed at the bottom of the main cylinder for extracting the test solution in the circular groove through the puncture assembly.

[0013] Preferably, the storage unit includes a tank placed in a circular groove, an inner tank slidably inserted inside the tank, and several strip grooves distributed along its axis on the outer side of the inner tank. An upper cover plate is provided on the inner tank by means of a threaded connection, and the outer side of the upper cover plate extends to the upper end of the tank, while sealing the through opening between the inner tank and the tank body. A structural groove is also provided on the inner side wall of the tank.

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

[0015] Preferably, the puncture assembly includes several puncture grooves on the support plate that correspond one-to-one with the through grooves. Several puncture shafts corresponding to the puncture grooves are provided on the bottom wall of the lower chamber. The top of the puncture shaft extends through the puncture groove into the corresponding circular groove and contacts the bottom of the sealing block. A conveying groove extending to the top of one side is provided in the puncture shaft. A bending groove corresponding to and communicating with the conveying groove is provided in the sealing block. The other end of the bending groove passes through the outside of the sealing block and communicates with the inside of the tank. A pipetting groove is provided on the outside of both the puncture shaft and the sealing block.

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

[0017] The lower chamber is also equipped with several fluid storage boxes that correspond one-to-one with the puncture shaft. One side of each fluid storage box has an infusion tube that extends into the puncture shaft and is connected to the delivery groove.

[0018] A push plate slides inside the liquid storage box, and push screws that are threadedly connected to the push plate are rotatably installed on both sides of the liquid storage box.

[0019] Preferably, the adsorption component includes two clips symmetrically arranged in a circular groove, with the test strip held between the clips.

[0020] Preferably, the limiting component includes a sealing sleeve installed on the upper end of the main cylinder. The inner side of the sealing sleeve is provided with several collars corresponding to the circular grooves. An upper plate that slides inside the collar and contacts the corresponding upper cover plate is connected to the sealing sleeve by a reset push spring.

[0021] Preferably, the bottom wall of the circular groove is provided with an annular ring, 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 bottom wall of the circular groove, and an active push spring is provided between the bottom wall of the annular groove and the annular ring.

[0022] Preferably, the vibration assembly includes a support plate disposed on the upper end of the support plate, a high-frequency vibration motor mounted on the support plate, an extension frame disposed on the outside of the vibration motor, and a transmission ring sleeved on the outside of the corresponding tank body at the ends of several extension sections of the extension frame.

[0023] In addition, the present invention also provides a method for detecting and identifying microorganisms, comprising the following steps:

[0024] S1, Sample processing: The collected sample is placed in the storage unit, which is then placed in the circular groove. Extraction reagent is then injected into the sample through the puncture assembly to separate the microbial DNA and RNA nucleic acid to obtain a mixed solution.

[0025] S2, Nucleic Acid Extraction: The mixed solution then enters the circular trough, where the adsorption component adsorbs and extracts the nucleic acid, while the remaining mixed solution is discharged from the trough by the puncture component.

[0026] S3, Nucleic Acid Solution Transfer: New extraction reagent is injected again into the circular groove through the puncture component to mix the nucleic acid attached to the adsorption component, resulting in a highly pure nucleic acid solution. The nucleic acid solution is then transported to the nanopore analyzer via the puncture component.

[0027] S4, Nanopore sequencer sequencing: Using a nanopore sequencer, nucleic acid solutions are sequenced. The nanopore sequencer starts a sequencing and analysis process. After obtaining enough high-quality sequencing data to determine the quality of the data, a test report is issued based on the actual situation. Sequencing continues as needed, and test reports are issued for verification and comparison under different sequencing data volumes.

[0028] In summary, this application includes at least one of the following beneficial technical effects:

[0029] I. This invention, through the cooperation of a storage unit, a puncture component, and an adsorption component, can efficiently separate microbial DNA and RNA nucleic acids during microbial sample detection, and utilize the adsorption component to extract nucleic acids a second time, accurately improving the purity of nucleic acid samples and solving the problems of low purity and many impurities in traditional methods.

[0030] Second, this invention utilizes the on-demand sequencing and analysis capabilities of the nanopore analyzer, combined with clear data quality and positive judgment criteria, to flexibly and quickly generate test reports after obtaining sufficient high-quality sequencing data, without having to complete the entire sequencing process, thus greatly shortening the testing cycle and improving testing efficiency.

[0031] Third, this invention automates operations such as moving the support plate and transporting extraction reagents through the coordinated operation of various transmission components in the drive unit, reducing manual intervention and human error. At the same time, the drive unit, in conjunction with other components, further improves the automation level and accuracy of the entire microbial detection and identification process. Attached Figure Description

[0032] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0033] Figure 1 This is a schematic diagram of the main body of the present invention.

[0034] Figure 2 This is a cross-sectional view of the main body of the present invention.

[0035] Figure 3 This is a cross-sectional view of the storage unit of the present invention.

[0036] Figure 4 This is the present invention. Figure 3 Enlarged view of part of the structure at point A in the middle.

[0037] Figure 5 This is a cross-sectional view of the puncture assembly of the present invention.

[0038] Figure 6 This is a schematic diagram of the adsorption component of the present invention.

[0039] Figure 7 This is the present invention. Figure 6 Enlarged view of part of the structure at point B.

[0040] Figure 8 This is a cross-sectional view of the limiting component and the vibration component of the present invention.

[0041] Figure 9 This is a schematic diagram of the structure of the driving unit of the present invention.

[0042] Figure 10 This is a diagram showing the combination of the arc-shaped magnetic plate and the passive magnetic plate of the present invention.

[0043] Figure 11 This is a diagram showing the assembly of the vertical axis, horizontal axis, drive shaft, and reciprocating lead screw of this invention.

[0044] In the diagram, 1. Main cylinder; 10. Support plate; 11. Circular groove; 12. Nanopore tester; 2. Storage unit; 20. Tank body; 21. Inner tank; 22. Strip groove; 23. Top cover plate; 24. Structural groove; 25. Sealing block; 3. Puncture assembly; 30. Puncture groove; 31. Puncture shaft; 32. Conveying groove; 33. Bending groove; 34. Pipette; 35. Infusion tank; 36. Bending tube; 37. Storage box; 38. Infusion tube; 39. Push plate; 310. 4. Drive screw; 5. Adsorption assembly; 6. Clip; 7. Test paper; 8. Limiting assembly; 9. Sealing sleeve; 10. Collar; 11. Upper plate; 2. Annular ring; 3. Limiting shaft; 4. Vibration assembly; 50. Vibration motor; 61. Transmission ring; 72. Drive unit; 73. Drive cavity; 74. Vertical groove; 75. Vertical shaft; 76. Horizontal shaft; 77. Drive shaft; 88. Arc groove; 99. Reciprocating screw; 10. Arc magnetic plate; 11. Passive magnetic plate. Detailed Implementation

[0045] The following combination Figures 1 to 11 The embodiments of the present invention will be described in detail below.

[0046] This application discloses a microbial detection and identification device and method. Applied to the detection and identification of microbial samples, this application achieves efficient separation of microbial DNA and RNA nucleic acids through innovative structural design, and accurately extracts nucleic acid samples, solving the problems of cumbersome procedures and low extraction purity in traditional methods. Furthermore, this application can also achieve simultaneous sequencing and analysis using a nanopore analyzer, flexibly generating test reports based on strict data quality and positive judgment standards. Simultaneously, the application utilizes a drive unit to automate the device's operation, significantly improving detection and identification efficiency and accuracy, providing a novel technical solution for the field of microbial detection.

[0047] Example 1: Refer to Figure 1 , Figure 2 and Figure 3 As shown, the device includes a main cylinder 1, a support plate 10, a circular groove 11, a storage unit 2, a puncture assembly 3, an adsorption assembly 4, a limiting assembly 5, a vibration assembly 6, and a nanopore tester 12. The support plate 10 is slidably arranged in the middle of the main cylinder 1, and the support plate 10 divides the main cylinder 1 into an upper chamber and a lower chamber. Several circular grooves 11 are opened on the support plate 10. The storage unit 2 for storing extracts is placed in the circular grooves 11, and the puncture assembly 3 is arranged in the lower chamber. One side of the puncture assembly 3 extends into the circular grooves 11 and the storage unit 2 and is used to replenish the extraction reagent inside them.

[0048] Furthermore, the circular groove 11 is also equipped with an adsorption component 4, which is used to adsorb the DNA and RNA of microorganisms in the extraction reagent.

[0049] The upper end of the main cylinder 1 is provided with a limiting component 5 for limiting and sealing the storage unit 2.

[0050] A vibration assembly 6 is installed on the support plate 10.

[0051] A nanopore tester 12, which is connected to the puncture assembly 3, is installed at the bottom of the main cylinder 1 and is used to extract the test solution in the circular groove 11 through the puncture assembly 3.

[0052] In addition, the present invention also provides a method for detecting and identifying microorganisms, comprising the following steps:

[0053] S1, Sample processing: The collected sample is placed in storage unit 2, then storage unit 2 is placed in circular groove 11, and then extraction reagent is injected into the sample through puncture component 3 to separate microbial DNA and RNA nucleic acid to obtain a mixed solution.

[0054] S2, Nucleic acid extraction: The mixed solution then enters the circular groove 11, where the adsorption component 4 adsorbs and extracts the nucleic acid, and the remaining mixed solution is discharged from the circular groove 11 by the puncture component 3.

[0055] S3, Nucleic acid solution transfer: New extraction reagent is injected again into the circular groove 11 through the puncture component 3 to mix the nucleic acid adsorbed on the adsorption component 4, resulting in a high-purity nucleic acid solution. The nucleic acid solution is then transported to the nanopore tester 12 through the puncture component 3.

[0056] S4, Nanopore sequencer sequencing: Using the nanopore sequencer 12, the nucleic acid solution is sequenced. The nanopore sequencing process begins with sequencing and analysis simultaneously. After obtaining sufficient high-quality sequencing data for data assessment, a test report is issued based on the actual situation. Sequencing continues as needed, and test reports are issued for verification and comparison under different sequencing data volumes.

[0057] Reference Figure 3 and Figure 4 As shown, a storage unit 2 for storing extracts 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 sealing 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. Several strip grooves 22 distributed along its axis are opened on the outer side of the inner tank 21. The 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, while sealing the through opening between the inner tank 21 and the tank body 20.

[0058] The inner wall of the tank body 20 is also provided with a structural groove 24.

[0059] The bottom of the tank body 20 is provided with a through groove, and the bottom of the inner tank 21 is provided with a sealing block 25 extending into the through groove.

[0060] In actual use, the swabs or test strips used to collect microorganisms (for ease of explanation, swabs or test strips are collectively referred to as collected materials) are placed in the inner container 21. Then, the top cover 23 is threaded into the top of the inner container 21. The inner container 21 is then installed in the container body 20, and the container body 20 is placed in the circular groove 11. After the inner container 21 is inserted into the container body 20, the sealing block 25 will simultaneously seal the through groove. At this time, the extraction reagent is injected into the container body 20. The extraction reagent dilutes the collected materials, and after the reaction occurs, the microbial nucleic acid in the collected materials will enter the extraction reagent.

[0061] Reference Figure 3 As shown, this is the puncture assembly 3 used to replenish the extraction reagent inside the storage unit 2; specifically, the puncture assembly 3 includes a puncture groove 30, a puncture shaft 31, a delivery groove 32, a bending groove 33, a pipetting groove 34, an infusion groove 35, a bending tube 36, a storage box 37, an infusion tube 38, a push plate 39, and a push screw 310. Several puncture grooves 30 are formed on the support plate 10 and correspond one-to-one with the through groove. 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 30. The piercing shaft 31 extends into the corresponding circular groove 11 and contacts the bottom of the sealing block 25. In actual operation, a sealing ring (known technology) can be installed on the bottom of the piercing shaft 31 and the sealing block 25 to prevent liquid leakage. This is a routine operation for those skilled in the art, so it will not be described in detail here. The piercing shaft 31 has a conveying groove 32 extending to its top on one side, and the sealing block 25 has a bent groove 33 with its bottom corresponding to and communicating with the conveying groove 32. The other end of the bent groove 33 passes through the outside of the sealing block 25 and communicates with the inside of the tank 20.

[0062] That is, when the support plate 10 drives the tank 20 to descend in the main cylinder 1, the support plate 10 is moving relative to the piercing shaft 31. Therefore, the piercing shaft 31 can enter the circular groove 11 through the piercing groove 30 and then contact the bottom of the sealing block 25. The bottom of the bending groove 33 will be connected to the top of the conveying groove 32.

[0063] Both the puncture shaft 31 and the sealing block 25 have a pipetting groove 34 on their outer sides.

[0064] The puncture shaft 31 is also provided with an infusion tank 35 with both ends extending to its outer side. A bent tube 36 is connected to the outer side of the puncture shaft 31, and one side of the bent tube 36 extends to the nanopore tester 12 and is connected to its test port.

[0065] The lower chamber is also equipped with several liquid storage boxes 37 that correspond one-to-one with the puncture shaft 31. One side of the liquid storage box 37 has an infusion tube 38 that extends into the interior of the puncture shaft 31 and is connected to the delivery groove 32.

[0066] A push plate 39 slides inside the liquid storage box 37, and push screws 310 that are threadedly connected to the push plate 39 are rotatably inserted on both sides of the liquid storage box 37.

[0067] In actual use, the push screw 310 is driven by external force to move the push plate 39 within the liquid storage box 37. The liquid storage box 37 is used to store the extraction reagent. The push plate 39 can drive the extraction reagent from the infusion tube 38 into the delivery tank 32. The extraction reagent then enters the bending tank 33 through the delivery tank 32 and is finally discharged into the tank 20. After that, the extraction reagent enters the inner tank 21 from the construction tank 24 and the strip tank 22 to come into contact with 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, which is the initial extraction.

[0068] Furthermore, several external connecting pipes that communicate with the liquid storage box 37 are inserted at the lower end of the main cylinder 1, allowing the operator to supply extraction reagents into the liquid storage box 37 through the external connecting pipes.

[0069] After the initial extraction is completed, the support plate 10 continues to descend, causing the puncture shaft 31 to continue to separate the inner tank 21 from the tank body 20 through the sealing block 25. This allows the liquid in the inner tank 21 to flow into the tank body 20 through the strip groove 22 and the structural groove 24. 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. 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 to the circular groove 11 through the structural groove 24. The extraction reagent in the tank body 20 will enter the circular groove 11 through the structural groove 24.

[0070] Since the extraction reagent at this time contains not only the nucleic acid of microorganisms, 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 34 outside the puncture shaft 31 is connected with the inside of the circular groove 11. The extraction reagent in the circular groove 11 will enter the pipette 34. Then the extraction reagent will enter the nanopore tester 12 through the pipette 34 and the bent tube 36. At this time, the nanopore tester 12 will not test the extraction reagent and will directly discharge it outside the nanopore tester 12.

[0071] Then the support plate 10 moves upward so that the top of the puncture shaft 31 is located in the circular groove 11. At this time, the liquid storage box 37 discharges the extraction reagent into the circular groove 11 again through the delivery groove 32, so that the extraction reagent extracts the nucleic acid in the adsorption component 4. The extracted nucleic acid is pure. After going through the above steps again, the extraction reagent can be discharged into the nanopore tester 12, and the nucleic acid in the extraction reagent can be tested by the nanopore tester 12.

[0072] The Nanopore Analyzer 12 can perform sequencing and analysis on the extraction reagents simultaneously, providing analysis results without completing the entire sequencing process. Once sufficient high-quality sequencing data has been obtained, a test report can be issued based on the actual situation. Sequencing can also continue as needed, and test reports can be issued for verification and comparison under different sequencing data volumes.

[0073] Data quality assessment criteria, examples are as follows:

[0074] (1) Ctr11 (external parameter sequence 1), the target is working and the number of reads is normal, such as reads < 10; (2) Ctr12 (external parameter sequence 2), 8 targets are working and the proportion is normal.

[0075] (3) Cleanreads account for >70%.

[0076] (4) More than 20,000 cleanreads were matched.

[0077] The criteria for interpreting a positive test result are shown below:

[0078] (1) The number of amplicon working units is greater than 3.

[0079] (2) The number of reads is consistent with the standard curve of the product design (color judgment).

[0080] (3) There is no cross-contamination or aerosol contamination signal interference between samples.

[0081] After the device has been tested, the cleaning operation can be carried out according to the following procedure: Remove the tank 20 from the circular groove 11, inject cleaning fluid into the storage box 37 through the external connecting pipe, push the lead screw 310 to drive the push plate 39, so that the cleaning fluid enters the delivery groove 32 of the puncture shaft 31 through the infusion pipe 38, and then flows into the tank 20 through the bending groove 33 of the sealing block 25 to rinse the nucleic acid impurities remaining on the inner wall of the tank 20, the structural groove 24 and the surface of the inner tank 21.

[0082] After rinsing, the waste liquid enters the infusion tank 35 of the puncture shaft 31 through the construction tank 24 and the transfer tank 34 at the bottom of the circular tank 11. It then flows into the nanopore tester 12 through the bent tube 36 and is finally discharged from the tester's discharge port.

[0083] At the same time, the vibration component 6 vibrates to enhance the flushing effect of the cleaning fluid on the inner wall of the circular groove and the residual dirt in the puncture groove 30, ensuring that there is no risk of cross-contamination of samples inside the device and preparing for the next test.

[0084] 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 clips 40 and test strips 41. The two clips 40 are symmetrically arranged inside the circular groove 11, and the test strips 41 are held between the clips 40.

[0085] The clips 40 at both ends can hold the test strip 41. When the test strip 41 first enters the circular groove 11, it contains a large amount of nucleic acid. Therefore, the test strip 41 can adsorb some of the nucleic acid in the extracting reagent. When the extracting reagent is discharged from the circular groove 11, the extracting reagent that enters the circular groove 11 a second time does not contain nucleic acid. Since the test strip 41 has already reacted once, when the extracting reagent that enters the circular groove 11 a second time comes into contact with the test strip 41, the nucleic acid in the test strip 41 will enter the extracting reagent.

[0086] In actual use, first clamp the corresponding test strip 41 onto the corresponding clip 40, and then install the storage unit 2 into the circular groove 11.

[0087] Reference Figure 4 and Figure 8 As shown, the upper end of the main cylinder 1 is provided with a limiting component 5 for limiting and sealing the storage unit 2; specifically, the limiting component 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 cylinder 1. Several collars 51 corresponding to the circular groove 11 are provided inside the sealing sleeve 50. The upper plate 52 that slides inside the collar 51 and contacts the corresponding upper cover plate 23 is connected to the sealing sleeve 50 by a reset push spring.

[0088] 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 cylinder 1. 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. When the inner tank 21 is driven by the piercing shaft 31 to move up and down in the tank body 20, the upper plate 52 will also move back and forth in the collar 51 and always abut against the inner tank 21 through the upper cover plate 23, so that when the piercing shaft 31 no longer abuts against the inner tank 21, the inner tank 21 can slide back into the tank body 20.

[0089] 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. A number of limiting shafts 54 that slide through the bottom of the tank body 20 are provided on one end of the annular ring 53. 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.

[0090] That is, after the tank 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 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 20 and the bottom wall of the circular groove 11 to prevent the bottom of the tank 20 from colliding with the adsorption component 4. The limiting shaft 54 ​​can limit the tank 20 to prevent the tank 20 from accidentally displacing or rotating on the annular ring 53.

[0091] 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. A support plate is set on the upper end of the support plate 10, and a 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 fitted with transfer rings 61 that are sleeved on the outside of the corresponding tank body 20. The transfer rings 61 are slidably connected to the tank body 20.

[0092] When the vibration motor 60 is running, it generates vibrations. These vibrations are transmitted to the tank 20 through the extension frame and the transfer ring 61, which promotes more thorough mixing and reaction between the extraction reagent and the extract. When the inner tank 21 moves upward and the extraction reagent flows into the tank 20 through the construction groove 24, the vibration can accelerate the flow rate of the solution. In addition, the vibration can also significantly improve the reaction efficiency of each step in the subsequent adsorption reaction between the extraction reagent and the test strip 41, and in the process of the extraction reagent entering the circular groove 11 a second time to extract the nucleic acid from the test strip 41.

[0093] Furthermore, when the tank 20 vibrates, the corresponding active push springs on its upper plate 52 and annular ring 53 can play a buffering and enhancing role, while the limiting shaft 54 ​​can play a limiting role on the tank 20 at this time.

[0094] Further explanation: The test strip 41 is based on a glass fiber membrane with a surface-modified silicon-based adsorption material. The silanol groups on its surface can physically adsorb nucleic acids through electrostatic interaction. When the mixed solution containing nucleic acids enters the designated circular groove or other structure for the first time, the vibration component 6 is activated, driving the relevant components to operate and ensuring that the test strip 41 is in full contact with the solution. Relying on the electrostatic adsorption capacity of the silanol groups, the nucleic acids in the solution are adsorbed and retained on the test strip 41. Impurities that may be present in the solution are not adsorbed by the test strip 41 and remain in the initial mixed solution.

[0095] After the initial 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. The extraction reagent, in conjunction with the force generated by the vibration, gradually breaks the electrostatic bond between the silanol group and the nucleic acid. Since the physical adsorption of the test strip 41 is a reversible process and it does not have any special chemical modification or specific binding groups, in this adsorption-desorption cycle, the nucleic acid adsorbed on the test strip 41 will be partially released into the injected extraction reagent solution.

[0096] The key role of this process is to allow the nucleic acid released from the test strip 41 into the solution to be as free as possible from the interference of impurities in the initial mixed solution. Although it cannot be guaranteed that the solution contains only nucleic acid, this secondary treatment can significantly reduce the impurity content and improve the accuracy of subsequent tests.

[0097] Example 2: Refer to Figure 9 , Figure 10 and Figure 11 As shown, based on Embodiment 1, in order to drive the support plate 10 and the support plate 10 to move, a drive cavity 70 is provided inside the main cylinder 1, and a drive unit 7 is provided inside the drive cavity 70; specifically, the drive unit 7 includes a drive cavity 70, a vertical groove 71, a vertical shaft 72, a horizontal shaft 73, a drive shaft 74, an arc-shaped groove 75, a reciprocating screw 76, an arc-shaped magnetic plate 77, and a passive magnetic plate 78. Several vertical grooves 71, corresponding one-to-one with the liquid storage box 37, are opened inside the main cylinder 1. The bottom of the vertical groove 71 communicates with the drive cavity 70, and a vertical shaft 72 rotates inside the vertical groove 71. Several drives 78 rotate through the inner wall of the main cylinder 1. A horizontal shaft 73 is located in the corresponding vertical groove 71, and the horizontal shaft 73 is connected to the vertical shaft 72 on one side of the vertical groove 71 through 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 push screw 310. The vertical shaft 72 can rotate in the vertical groove 71, and during the rotation of the vertical shaft 72, it can also drive the horizontal shaft 73 to rotate through gear transmission. When the horizontal shaft 73 rotates, it can drive the corresponding push screw 310 to rotate synchronously, so that the push screw 310 drives the corresponding push plate 39 to reciprocate within the liquid storage box 37.

[0098] A drive shaft 74 is also provided on the inner bottom wall of the drive cavity 70. The bottom of several vertical shafts 72 extends into the drive cavity 70 and is connected to each other by belt drive. The vertical shafts 72 on one side are connected to the drive shaft 74 by belt drive. That is, the vertical shafts 72 can rotate synchronously by gear drive. When the drive shaft 74 is driven by an external drive device (existing motor device), it can drive the vertical shafts 72 on one side to rotate by belt drive, so that all the vertical shafts 72 are driven to rotate.

[0099] 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 76 is rotated on the top wall of the arc-shaped groove 75, and an arc-shaped magnetic plate 77 is threadedly connected to the outside of the reciprocating screw 76 and slidably disposed in the arc-shaped groove 75. 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 76 extends into the drive cavity 70 and is connected to the drive shaft 74 by belt drive. During the rotation of the drive shaft 74, the reciprocating screw 76 can also be driven to rotate by belt drive. When the reciprocating screw 76 rotates, it can also drive the arc-shaped magnetic plate 77 to move up and down in the arc-shaped groove 75. When the arc-shaped magnetic plate 77 moves, it can also drive the passive magnetic plate 78 and the support plate 10 connected to it to move up and down in the main cylinder 1 by magnetic connection.

[0100] 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 invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered in all respects as exemplary and not restrictive.

[0101] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments 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 support plate (10) is slidably provided in the middle of the main cylinder (1), and the support plate (10) divides the main cylinder (1) into an upper chamber and a lower chamber. Several circular grooves (11) are provided on the support plate (10), and a storage unit (2) for storing extract is placed in the circular groove (11). 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 extract reagent inside. Furthermore, the circular groove (11) is also equipped with an adsorption component (4) for adsorbing the DNA and RNA of microorganisms in the extraction reagent; The upper end of the main cylinder (1) is provided with a limiting component (5) for limiting and sealing the storage unit (2); the support plate (10) is provided with a vibration component (6). A nanopore tester (12) connected to the puncture assembly (3) is installed at the bottom of the main cylinder (1) for extracting the test solution in the circular groove (11) through the puncture assembly (3); The storage unit (2) includes a tank (20) placed in a circular groove (11). An inner tank (21) is slidably inserted inside the tank (20). Several strip grooves (22) distributed along its axis are opened on the outer side of the inner tank (21). An upper cover plate (23) is provided on the inner tank (21) by means of threaded connection. The outer side of the upper cover plate (23) extends to the upper end of the tank (20) and seals the through opening between the inner tank (21) and the tank (20). A structural groove (24) is also opened on the inner side wall of the tank (20). The bottom of the tank body (20) is provided with a through groove, and the bottom of the inner tank (21) is provided with a sealing block (25) extending into the through groove. The puncture assembly (3) includes several puncture grooves (30) that are opened on the support plate (10) and correspond one-to-one with the through grooves. 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 (30) and extends into the corresponding circular groove (11) and contacts the bottom of the sealing block (25). A conveying groove (32) extending to the top of one side is opened in the puncture shaft (31). A bending groove (33) corresponding to and communicating with the conveying groove (32) is opened in the sealing block (25). The other end of the bending groove (33) passes through the outside of the sealing block (25) and communicates with the inside of the tank (20). A pipetting groove (34) is opened on the outside of both the puncture shaft (31) and the sealing block (25).

2. The microbial detection and identification device according to claim 1, characterized in that: The puncture shaft (31) is also provided with an infusion tank (35) with both ends extending to its outer side. A bent tube (36) is connected to the outside of the puncture shaft (31), and one side of the bent tube (36) extends to the nanopore tester (12) and is connected to its test port. The lower chamber is also equipped with several reservoir boxes (37) that correspond one-to-one with the puncture shaft (31). One side of the reservoir box (37) has an infusion tube (38) that extends into the puncture shaft (31) and is connected to the delivery groove (32). A push plate (39) slides inside the liquid storage box (37), and push screws (310) that are threadedly connected to the push plate (39) are rotatably installed on both sides of the liquid storage box (37).

3. The microbial detection and identification device according to claim 1, characterized in that: The adsorption assembly (4) includes two clips (40) symmetrically arranged in a circular groove (11), with a test paper (41) held between the clips (40).

4. The microbial detection and identification device according to claim 1, characterized in that: The limiting component (5) includes a sealing sleeve (50) installed on the upper end of the main cylinder (1). The inner side of the sealing sleeve (50) is provided with several collars (51) corresponding to the circular groove (11). The collars (51) have an upper plate (52) that slides in contact with the corresponding upper cover plate (23). The upper plate (52) and the sealing sleeve (50) are connected by a reset push spring.

5. The microbial detection and identification device according to claim 4, characterized in that: The inner bottom wall of the circular groove (11) is provided with an annular ring (53), 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 several limiting shafts (54) that slide 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).

6. The microbial detection and identification device according to claim 1, characterized in that: The vibration assembly (6) includes a support plate set on the upper end of the support plate (10), a high-frequency vibration motor (60) is installed on the support plate, an extension frame is set on the outside of the high-frequency vibration motor (60), and a transmission ring (61) sleeved on the outside of the corresponding tank body (20) is installed at the ends of several extension sections of the extension frame.

7. A method for detecting and identifying microorganisms, using a microorganism detection and identification device as described in any one of claims 1-6, characterized in that, The identification method includes 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). Then, the extraction reagent is injected into the sample through the puncture assembly (3) so that the microbial DNA and RNA nucleic acid are separated to obtain a mixed solution. S2, Nucleic acid extraction: The mixed solution then enters the circular groove (11), where the adsorption component (4) 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: New extraction reagent is injected into the circular groove (11) again through the puncture component (3) to mix the nucleic acid attached to the adsorption component (4) and obtain a nucleic acid solution with high purity. The nucleic acid solution is then transported to the nanopore tester (12) through the puncture component (3). S4, Nanopore sequencer sequencing: Using a nanopore sequencer (12), the nucleic acid solution is sequenced. The nanopore sequencer (12) starts the sequencing and analysis process. After obtaining enough high-quality sequencing data for data judgment, a test report is issued according to the actual situation. Sequencing continues according to actual needs, and test reports are issued under different sequencing data volumes for verification and comparison.

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