Microbiological automated detection carousel and device

Abstract

The application provides a microorganism automatic detection turntable and device, which comprises a top plate, a middle layer plate, a one-way conduction structure and a bottom plate which are fixed and connected in sequence from top to bottom, and the top plate is slidably connected with the middle layer plate; a sample chamber is arranged on the top plate, and a plurality of detection structures are arranged on the top plate in a circumferential distribution; the detection structure comprises a nucleic acid extraction chamber, a washing liquid chamber and a nucleic acid detection chamber, and the sample chamber is communicated with the nucleic acid extraction chamber; a plurality of bacterial interception structures are arranged on the middle layer plate in a circumferential distribution, the bacterial interception structure is used for intercepting bacteria in a to-be-detected solution, and waste liquid is discharged to the bottom plate through the one-way conduction structure; the one-way conduction structure is used for preventing the waste liquid discharged to the bottom plate from flowing back to the bacterial interception structure; and a plurality of waste liquid storage structures are arranged on the bottom plate in a circumferential distribution. The application realizes rapid and simple detection of foodborne pathogenic bacteria and reduces the dependence on large instruments and technical personnel.

Description

Technical Field

[0001] This invention relates to the field of microbial detection technology, and in particular to an automated microbial detection turntable and device. Background Technology

[0002] Foodborne pathogen contamination occurs at every stage of the food production, processing, storage, transportation, and consumption chain. Therefore, corresponding detection methods should be simple, rapid, and portable to meet the detection needs of different scenarios. However, existing methods for detecting foodborne pathogens mainly include culture methods, immunological methods (such as ELISA), and molecular biological methods (such as PCR). Culture methods are the gold standard for bacterial detection, with high accuracy and sensitivity, but are time-consuming; ELISA has a shorter detection time, better specificity, and is easy to achieve high throughput, but has lower sensitivity and a higher false positive rate; PCR also has a shorter detection time and higher sensitivity, but nucleic acid extraction is complex and requires expensive equipment and professional technicians.

[0003] Therefore, how to shorten detection time, simplify operation, and reduce reliance on large instruments and technical personnel while ensuring high sensitivity is an important issue that the industry urgently needs to address. Summary of the Invention

[0004] This invention provides an automated microbial detection turntable and device to address the shortcomings of existing foodborne pathogen detection methods, such as long detection time, low detection sensitivity, complex operation, and high dependence on large instruments and technicians. It achieves rapid and convenient detection of foodborne pathogens while maintaining sensitivity, and reduces dependence on large instruments and technicians.

[0005] This invention provides an automated microbial detection turntable, comprising a top plate and a middle plate, a unidirectional conduction structure, and a bottom plate that are fixedly connected from top to bottom, wherein the top plate and the middle plate are slidably connected.

[0006] The top plate is provided with a sample chamber, and the top plate is also provided with multiple detection structures distributed along the circumference;

[0007] The detection structure includes a nucleic acid extraction chamber, a washing solution chamber, and a nucleic acid detection chamber, wherein the sample chamber is connected to the nucleic acid extraction chamber;

[0008] The middle plate is provided with multiple bacterial retention structures distributed along the circumference. The bacterial retention structures are located below the nucleic acid extraction chamber. The bacterial retention structures are used to retain bacteria in the test solution and discharge waste liquid to the bottom plate through the one-way conduction structure.

[0009] The unidirectional flow structure is used to prevent waste liquid discharged to the bottom plate from flowing back to the bacterial retention structure;

[0010] The base plate is provided with multiple waste liquid storage structures distributed along the circumference, and the waste liquid storage structures are used to store waste liquid.

[0011] According to the present invention, an automated microbial detection turntable is provided, wherein a silicone oil layer is disposed between the top plate and the middle plate.

[0012] According to the present invention, an automated microbial detection turntable is provided, wherein the middle plate is provided with a groove that matches the top plate, and the bacterial retention structure is located at the bottom of the groove.

[0013] According to the present invention, an automated microbial detection turntable is provided, wherein the bacterial retention structure includes a retention chamber, an FTA card is disposed in the retention chamber, the retention chamber is located below the nucleic acid extraction chamber, and the bottom of the retention chamber is connected to the unidirectional conduction structure.

[0014] According to the present invention, an automated microbial detection turntable is provided, wherein the one-way conduction structure includes a thin film, and the thin film is provided with a plurality of one-way valve holes distributed along the circumference. The one-way valve holes are located between the bacterial interception structure and the waste liquid storage structure, and the one-way valve holes correspond one-to-one with the bacterial interception structure.

[0015] According to the present invention, an automated microbial detection turntable is provided, wherein the waste liquid storage structure includes a flow chamber and a waste liquid chamber, the flow chamber being connected to the waste liquid chamber and the flow chamber being located below the bacterial retention structure.

[0016] According to the present invention, in an automated microbial detection turntable, the circumferences of the nucleic acid extraction chamber, the washing solution chamber, and the nucleic acid detection chamber coincide with the center of the circumference of the bacterial retention chamber, are the same size, and are parallel to each other.

[0017] The present invention also provides an automated microbial detection device, comprising a detection chamber, a cover connected to the detection chamber, a rotation drive, a heating element, a pressing assembly, and an automated microbial detection turntable. The rotation drive is fixedly installed inside the chamber, the automated microbial detection turntable is installed on the detection chamber, the rotation drive is connected to the automated microbial detection turntable, the heating element is fixedly installed on the detection chamber and is used to heat the automated microbial detection turntable, and the pressing assembly is installed on the cover and is located above the automated microbial detection turntable.

[0018] According to the present invention, an automated microbial detection turntable is provided, wherein the clamping assembly includes a clamping drive and a pressure plate, the clamping drive is fixedly installed on the box cover, and the pressure plate is connected to the clamping drive.

[0019] According to the present invention, an automated microbial detection turntable is provided, wherein a camera is also provided on the lid, and the camera is located between the pressure plate and the lid.

[0020] According to the automated microbial detection turntable and device provided by the present invention, the bacterial solution to be tested is first added to the sample chamber, and the relative positions of the top plate and the middle plate are adjusted so that the bacterial retention structure is located below the nucleic acid extraction chamber. This allows the bacterial solution to flow into the nucleic acid extraction chamber and then into the bacterial retention structure. Then, the automated microbial detection turntable is rotated at high speed by an external device, causing the bacterial solution to be driven by centrifugal force. The bacteria in the bacterial solution are retained in the bacterial retention structure, while the waste liquid in the bacterial solution is discharged into the waste liquid storage structure for storage through a one-way flow structure. The bacterial retention structure is then heated, causing the bacteria to lyse and release their nucleic acid substances. The top plate is then rotated so that the washing chamber rotates above the bacterial retention structure. After standing for a certain period, the automated microbial detection turntable is rotated at high speed again, causing the waste liquid to flow through the bacterial retention structure and the one-way flow structure to the waste liquid storage structure for storage. The top plate is then rotated again, positioning the nucleic acid detection chamber above the bacterial trapping structure. Simultaneously, the chamber is heated, and recombinase-mediated isothermal amplification (RTA) technology is used to amplify the target microbial nucleic acid, causing a fluorescence change in the solution. An image of the nucleic acid detection chamber is acquired every 10 seconds, and its normalized G / (R+G+B) value is analyzed. The threshold time for fluorescence appearance is analyzed based on the dynamic normalized G / (R+G+B) value, and the bacterial content in the sample is calculated using a pre-calibrated threshold time-bacterial concentration curve. This integration of sample pretreatment, nucleic acid extraction, nucleic acid purification, and nucleic acid detection enables rapid and convenient detection of foodborne pathogens while maintaining sensitivity, reducing reliance on large instruments and technical personnel. Attached Figure Description

[0021] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0022] Figure 1 This is a schematic diagram of the structure of the automated microbial detection turntable provided by the present invention;

[0023] Figure 2 This is a schematic diagram of the detection structure of the automated microbial detection turntable provided by the present invention;

[0024] Figure 3 This is one of the structural schematic diagrams of the automated microbial detection device provided by the present invention;

[0025] Figure 4 This is the second schematic diagram of the structure of the automated microbial detection device provided by the present invention;

[0026] Figure label:

[0027] 1: Top slab; 2: Middle slab; 3: Unidirectional ventilation structure;

[0028] 4: Base plate; 5: Inspection box body; 6: Box lid;

[0029] 7: Heating element; 8: Clamping assembly; 11: Sample chamber;

[0030] 12: Detection structure; 13: Nucleic acid extraction chamber; 14: Washing solution chamber;

[0031] 15: Nucleic acid detection chamber; 21: Bacterial retention structure; 22: Groove;

[0032] 23: FTA card; 31: Membrane; 32: One-way valve port;

[0033] 41: Waste liquid storage structure; 42: Flow chamber; 43: Waste liquid chamber;

[0034] 81: Clamping drive component; 82: Pressure plate. Detailed Implementation

[0035] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0036] The following is combined Figures 1 to 4 This invention describes an automated microbial detection turntable and apparatus.

[0037] As attached Figure 1 and attached Figure 2 As shown, the automated microbial detection turntable includes a top plate 1 and a middle plate 2, a unidirectional conduction structure 3, and a bottom plate 4, which are fixedly connected from top to bottom. The top plate 1 and the middle plate 2 are slidably connected.

[0038] Specifically, the top plate 1 has a sample chamber 11 and multiple detection structures 12 arranged circumferentially on it. Each detection structure 12 includes a nucleic acid extraction chamber 13, a washing chamber 14, and a nucleic acid detection chamber 15. The sample chamber 11 is connected to the nucleic acid extraction chamber 13. The middle plate 2 has multiple bacterial retention structures 21 arranged circumferentially below the nucleic acid extraction chamber 13. These structures are used to retain bacteria in the test solution and discharge waste liquid through a one-way flow structure 3 to the bottom plate 4. The one-way flow structure 3 prevents the waste liquid discharged to the bottom plate 4 from flowing back to the bacterial retention structures 21. The bottom plate 4 has multiple waste liquid storage structures 41 arranged circumferentially for storing waste liquid.

[0039] In operation, the bacterial solution to be tested is first added to the sample chamber 11, and the relative positions of the top plate 1 and the middle plate 2 are adjusted so that the bacterial retention structure 21 is located below the nucleic acid extraction chamber 13. This allows the bacterial solution to flow into the nucleic acid extraction chamber 13 and then into the bacterial retention structure 21. Then, an external device rotates the automated microbial detection turntable at high speed, driving the bacterial solution with centrifugal force. The bacteria in the bacterial solution are retained in the bacterial retention structure 21, while the waste liquid is discharged into the waste liquid storage structure through the one-way flow structure 3 for storage. The bacterial retention structure 21 is then heated, causing the bacteria to lyse and release their nucleic acid. The top plate 1 is then rotated so that the washing chamber 14 rotates above the bacterial retention structure 21. After a certain period of settling, the automated microbial detection turntable is rotated at high speed again, allowing the waste liquid to flow through the bacterial retention structure 21 and the one-way flow structure 3 to the waste liquid storage structure 41 for storage. Then, the top plate 1 is rotated again, positioning the nucleic acid detection chamber 15 above the bacterial trapping structure 21. Simultaneously, the nucleic acid detection chamber 15 is heated, and recombinase-mediated isothermal amplification technology is used to amplify the target microbial nucleic acid, causing a fluorescence change in the solution. An image of the nucleic acid detection chamber 15 is acquired every 10 seconds, and its normalized G / (R+G+B) value is analyzed. The threshold time for fluorescence appearance is analyzed based on the dynamic normalized G / (R+G+B) value, and the bacterial content in the sample is calculated according to a pre-calibrated threshold time-bacterial concentration curve. This integration of sample pretreatment, nucleic acid extraction, nucleic acid purification, and nucleic acid detection steps enables rapid and convenient detection of foodborne pathogens while maintaining sensitivity, reducing reliance on large instruments and technical personnel.

[0040] Among them, as attached Figure 1 and attached Figure 2As shown, the circumferences of the nucleic acid extraction chamber 13, washing chamber 14, and nucleic acid detection chamber 15 coincide with the center of the circumference of the bacterial retention chamber, are the same size, and are parallel to each other. The bacterial retention chambers correspond one-to-one with the nucleic acid extraction chamber 13. In use, by rotating the top plate 1, the nucleic acid extraction chamber 13, washing chamber 14, and nucleic acid detection chamber 15 are sequentially rotated above the bacterial retention chamber, solving the technical problem that traditional microbial detection technologies cannot effectively integrate sample pretreatment, nucleic acid extraction, nucleic acid purification, and nucleic acid detection steps.

[0041] Furthermore, a silicone oil layer (not shown in the figure) is provided between the top plate 1 and the middle plate 2. During use, by applying silicone oil between the top plate 1 and the middle plate 2, the top plate 1 can rotate relative to the middle plate 2 while ensuring a tight seal between them. This rotation of the top plate 1 allows the nucleic acid extraction chamber 13, the washing chamber 14, and the nucleic acid detection chamber 15 to sequentially rotate above the bacterial retention chamber, effectively integrating the steps of bacterial retention, nucleic acid purification, and nucleic acid amplification. This enables rapid and convenient bacterial detection, shortening the detection time.

[0042] Among them, as attached Figure 1 As shown, the middle plate 2 has a groove 22 that matches the top plate 1, and the bacteria trapping structure 21 is located at the bottom of the groove 22. In use, the top plate 1 is snapped into the groove 22. The silicone oil layer between the top plate 1 and the middle plate 2 keeps the connection between the top plate 1 and the groove 22 sealed, and the top plate 1 can also rotate relative to the groove 22.

[0043] Further details are attached. Figure 1 As shown, the bacterial retention structure 21 includes a retention chamber containing an FTA card 23. The retention chamber is located below the nucleic acid extraction chamber 13, and its bottom is connected to the unidirectional flow structure 3. In use, the bacterial solution to be tested is first added to the sample chamber 11. The bacterial solution flows into the nucleic acid extraction chamber 13 and then into the retention chamber. Waste liquid in the bacterial solution then flows through the FTA card 23 and the unidirectional flow structure to the waste liquid storage structure 41. The bacteria in the bacterial solution are enriched on the FTA card 23. The FTA card 23 is then dried. The bacteria on the FTA card 23 are lysed by a strong denaturing agent chelated on the fiber matrix, releasing their nucleic acid material. The nucleic acid material is adsorbed onto the FTA card 23, facilitating subsequent purification and amplification of the nucleic acid material.

[0044] Further details are attached. Figure 1As shown, the one-way flow structure 3 includes a membrane 31 with multiple circumferentially distributed one-way valve holes 32. These one-way valve holes 32 are located between the bacterial retention structure 21 and the waste liquid storage structure 41, with each one-to-one correspondence between the two. During use, waste liquid from the bacterial retention structure 21 flows through the one-way valve holes 32 into the waste liquid storage structure 41 for storage. Because the one-way valve holes 32 are one-way, waste liquid can only flow from the bacterial retention structure 21 to the waste liquid storage structure 41, and cannot flow from the waste liquid storage structure 41 back into the bacterial retention structure 21. This effectively prevents waste liquid backflow and ensures that bacterial detection can proceed normally.

[0045] In an optional embodiment of the present invention, the film 31 is, for example, a PDMS film 31. However, it should be understood that the film 31 can also be any other suitable type of film 31.

[0046] Further details are attached. Figure 1 As shown, the waste liquid storage structure 41 includes a flow chamber 42 and a waste liquid chamber 43, which are connected. The flow chamber 42 is located below the bacterial retention structure 21. During use, the waste liquid at the bacterial retention structure 21 flows through the unidirectional flow structure 3 into the flow chamber, and then into the waste liquid chamber 43 for storage. After bacterial detection, the waste liquid can be treated, reducing the waste liquid treatment steps during bacterial detection, further simplifying the bacterial detection process, enabling rapid and convenient bacterial detection, and shortening the detection time.

[0047] The waste liquid chamber 43 is equipped with an absorbent pad (not shown in the figure). During use, the absorbent pad can absorb and fix the waste liquid.

[0048] On the other hand, as attached Figure 3 and attached Figure 4 As shown, the present invention also provides an automated microbial detection device, including a detection chamber 5, a cover 6 connected to the detection chamber 5, a rotation drive, a heating element 7, and a clamping assembly 8. The rotation drive is fixedly installed inside the detection chamber 5, and the automated microbial detection turntable is installed on the detection chamber 5. The rotation drive is connected to the automated microbial detection turntable. The heating element 7 is fixedly installed on the detection chamber 5 and is used to heat the automated microbial detection turntable. The clamping assembly 8 is installed on the cover 6 and is located above the automated microbial detection turntable.

[0049] In use, open the lid 6, add the bacterial solution to be tested into the sample chamber 11, and then close the lid 6. The drive mechanism rotates the automated microbial detection turntable at high speed, causing the bacterial solution to be centrifugally driven. Bacteria in the solution are trapped in the bacterial trapping structure 21, while waste liquid is discharged into the waste liquid storage structure 41 through the one-way flow structure 3. The heating element 7 then heats the bacterial trapping structure 21, causing the bacteria to lyse and release their nucleic acid. The pressing assembly 8 then moves down to press against the top plate 1. The drive mechanism rotates the automated microbial detection turntable, causing the top plate 1 to rotate relative to the middle plate 2, positioning the washing chamber 14 above the bacterial trapping structure 21. After a certain period of settling, the pressing assembly 8 moves up to disengage from the top plate 1. The drive mechanism rotates the automated microbial detection turntable at high speed again, allowing the waste liquid to flow through the bacterial trapping structure 21 and the one-way flow structure 3 to the waste liquid storage structure 41 for storage. Then, the clamping component 8 moves down again to clamp the top plate 1. The rotating drive component drives the microbial automated detection turntable to rotate, causing the nucleic acid detection chamber 15 to rotate above the bacterial retention structure 21. Then, the heating component 7 heats the nucleic acid detection chamber 15, causing the solution to change fluorescence. The bacterial concentration can then be calculated. This achieves automated bacterial detection, integrating sample pretreatment, nucleic acid extraction, nucleic acid purification, and nucleic acid detection into one process. It enables rapid and convenient detection of foodborne pathogens while maintaining sensitivity, reducing reliance on large instruments and technical personnel.

[0050] In an optional embodiment of the invention, the rotation drive is, for example, a stepper motor. However, it should be understood that any other suitable drive can also be used as the rotation drive.

[0051] In an optional embodiment of the invention, the heating element 7 is, for example, a heating coil. However, it should be understood that the heating coil can also be any other suitable heating element.

[0052] Further details are attached. Figure 3 and attached Figure 4 As shown, the clamping assembly 8 includes a clamping drive 81 and a pressure plate 82. The clamping drive 81 is fixedly mounted on the cover 6, and the pressure plate 82 is connected to the clamping drive 81. In use, the clamping drive 81 drives the pressure plate 82 to move downward, so that the pressure plate 82 abuts against the top plate 1, thereby clamping the top plate 1; the clamping drive 81 drives the pressure plate 82 to move upward, so that the pressure plate 82 disengages from the top plate 1, thereby allowing the top plate 1 to rotate relative to the middle plate 2.

[0053] In an optional embodiment of the invention, the clamping drive 81 is, for example, an electrically controlled lead screw. However, it should be understood that the clamping drive 81 can also be any other suitable drive.

[0054] In an optional embodiment of the present invention, the pressure plate 82 is, for example, a transparent pressure plate 82.

[0055] Furthermore, a camera (not shown in the figure) is also installed on the lid 6, located between the pressure plate 82 and the lid 6. During use, when bacteria react with the solution in the nucleic acid detection chamber 15, the camera takes a picture of the nucleic acid detection chamber 15 every 10 seconds and transmits the captured image to the corresponding processor for processing.

[0056] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. An automated microbial detection turntable, characterized in that, It includes a top plate and a middle plate, a unidirectional conduction structure and a bottom plate that are fixedly connected from top to bottom, wherein the top plate and the middle plate are slidably connected. The top plate is provided with a sample chamber, and the top plate is also provided with multiple detection structures distributed along the circumference; The detection structure includes a nucleic acid extraction chamber, a washing solution chamber, and a nucleic acid detection chamber, wherein the sample chamber is connected to the nucleic acid extraction chamber; The middle plate is provided with multiple bacterial retention structures distributed along the circumference. The bacterial retention structures are located below the nucleic acid extraction chamber. The bacterial retention structures are used to retain bacteria in the test solution and discharge waste liquid to the bottom plate through the one-way conduction structure. The unidirectional flow structure is used to prevent waste liquid discharged to the bottom plate from flowing back to the bacterial retention structure; The base plate is provided with multiple waste liquid storage structures distributed along the circumference, and the waste liquid storage structures are used to store waste liquid.

2. The automated microbial detection turntable according to claim 1, characterized in that, A silicone oil layer is provided between the top plate and the middle plate.

3. The automated microbial detection turntable according to claim 2, characterized in that, The middle layer plate is provided with a groove that matches the top plate, and the bacterial trapping structure is located at the bottom of the groove.

4. The automated microbial detection rotary table according to any one of claims 1-3, characterized in that, The bacterial retention structure includes a retention chamber, in which an FTA card is disposed. The retention chamber is located below the nucleic acid extraction chamber, and the bottom of the retention chamber is connected to the unidirectional conduction structure.

5. The automated microbial detection rotary table according to any one of claims 1-3, characterized in that, The one-way conduction structure includes a membrane with a plurality of one-way valve holes distributed circumferentially on the membrane. The one-way valve holes are located between the bacterial retention structure and the waste liquid storage structure, and each one-way valve hole corresponds to a bacterial retention structure.

6. The automated microbial detection rotary table according to any one of claims 1-3, characterized in that, The waste liquid storage structure includes a flow chamber and a waste liquid chamber, the flow chamber being connected to the waste liquid chamber and located below the bacterial retention structure.

7. The automated microbial detection rotary table according to any one of claims 1-3, characterized in that, The circumferences of the nucleic acid extraction chamber, the washing solution chamber, and the nucleic acid detection chamber coincide with the center of the circumference of the bacterial retention structure, are the same size, and are parallel to each other.

8. An automated microbial detection device, characterized in that, The device includes a detection chamber, a cover connected to the detection chamber, a rotation drive, a heating element, a clamping assembly, and an automated microbial detection turntable as described in any one of claims 1-7. The rotation drive is fixedly installed inside the chamber, the automated microbial detection turntable is installed on the detection chamber, the rotation drive is connected to the automated microbial detection turntable, the heating element is fixedly installed on the detection chamber and is used to heat the automated microbial detection turntable, and the clamping assembly is installed on the cover and is located above the automated microbial detection turntable.

9. The automated microbial detection device according to claim 8, characterized in that, The clamping assembly includes a clamping drive and a pressure plate. The clamping drive is fixedly installed on the box cover, and the pressure plate is connected to the clamping drive.

10. The automated microbial detection device according to claim 9, characterized in that, A camera is also installed on the lid of the box, and the camera is located between the pressure plate and the lid of the box.

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