A tube-based bacterial / cellular detection device

Abstract

The utility model provides a kind of one tube bacterial / cell detection device, including test tube body, sealing cover, filter membrane and three-way valve, the test tube body has upper opening, the upper chamber being communicated with upper opening, the transition chamber being located below upper chamber and being communicated with upper chamber and the reaction chamber and waste liquid chamber being located below transition chamber and being communicated with transition chamber respectively;The sealing cover is used to seal cover and be closed in the upper opening of test tube body;The filter membrane is arranged in upper chamber;The three-way valve is arranged in transition chamber to control the on-off between upper chamber, reaction chamber and waste liquid chamber.Bacterial / cell collection, lysis to detection whole process is completed in test tube body, realizes one tube operation;Eliminate the step of sample transfer between multiple devices, compared with traditional mode has lower contamination rate and more simple operation, to improve the accuracy, convenience and reliability of detection.

Description

Technical Field

[0001] This utility model relates to the field of rapid biological sample analysis technology, specifically to a single-tube bacterial / cell detection device; in particular, it is an integrated device for collecting, lysing, and detecting the concentration of microorganisms / cells based on adenosine triphosphate (ATP) bioluminescence method. Background Technology

[0002] Rapid quantitative detection of bacteria and cells is of great value in clinical diagnostics, drug development, and biosafety monitoring. The ATP bioluminescence method is widely used for bacterial contamination assessment and cell viability detection due to its high sensitivity (detecting as few as one bacterium / cell) and universality (all living cells contain ATP). Its principle is that ATP reacts with luciferin under the catalysis of luciferase to emit fluorescence (wavelength 560 nm), and the fluorescence intensity is directly proportional to the number of living cells.

[0003] However, traditional ATP assays typically involve multiple separation steps:

[0004] Step 1. Sample filtration / concentration: The liquid to be tested is filtered through an independent filter membrane device to retain bacteria / cells on the filter membrane.

[0005] Step 2. Transfer and Lysis: Carefully remove the filter membrane containing bacteria / cells and transfer it to another reaction tube or container. Add lysis buffer to rupture the bacteria / cells and release ATP. This transfer step is cumbersome and easily contaminated or lost due to human error, affecting the accuracy of the test results.

[0006] Step 3. Reagent addition and detection: Add a detection solution containing luciferase and luciferin to the lysed sample, mix well, and transfer the mixture to a dedicated transparent detection tube or microplate. Use a fluorescence detector to read the luminescence value.

[0007] The above-mentioned ATP detection involves filtering bacteria or cells, then transferring the filter membrane to a lysis tube, and finally transferring the lysed liquid to a detection tube. This process involves multiple opening and closing of the cap, pipetting, or membrane transfer operations, which is not only cumbersome but also highly susceptible to sample contamination, bacterial / cell loss, or operational errors.

[0008] Therefore, there is an urgent need to develop an integrated device that truly achieves a single tube for the entire process, eliminates the need for sample transfer, is easy to operate, and effectively avoids contamination, in order to solve the problems of cumbersome steps, easy contamination, and low efficiency in existing technologies, and improve the accuracy, convenience, and reliability of detection. Utility Model Content

[0009] To address the aforementioned problems, this invention provides a single-tube bacterial / cell detection device.

[0010] To achieve the above objectives, the technical solution provided by this utility model is as follows:

[0011] A single-tube bacterial / cell detection device includes a test tube body, a sealing cap, a filter membrane, and a three-way valve. The test tube body has an upper opening, an upper chamber communicating with the upper opening, a transition chamber located below and communicating with the upper chamber, and a reaction chamber and a waste liquid chamber located below and communicating with the transition chamber. The sealing cap is used to seal the upper opening of the test tube body. The filter membrane is disposed in the upper chamber. The three-way valve is disposed in the transition chamber to control the flow between the upper chamber, the reaction chamber, and the waste liquid chamber.

[0012] Furthermore, the three-way valve is a manual valve, and the three-way valve is connected to a knob, which is externally located on the test tube body.

[0013] Furthermore, the knob is provided with a marking pattern.

[0014] Furthermore, the test tube body is made of transparent silicone.

[0015] Furthermore, the three-way valve is a remotely controllable electric valve.

[0016] Furthermore, the three-way valve is an L-type three-way ball valve.

[0017] Furthermore, the upper opening of the test tube body is connected to the sealing cap by a thread, forming a sealing fit.

[0018] Furthermore, the reaction chamber and the waste liquid chamber are distributed on the left and right sides.

[0019] Furthermore, the bottom of the test tube has a high-transmittance zone, which is part of the reaction chamber.

[0020] Furthermore, the bottom of the test tube is provided with a flat-bottomed upright section, and the high-transparency zone is located inside the upright section.

[0021] Furthermore, the upper chamber is also provided with a pre-filter membrane with a pore size larger than that of the filter membrane.

[0022] The technical solution provided by this utility model has the following beneficial effects:

[0023] The entire process from collection and lysis of bacteria / cells (i.e., bacteria or cells) to detection is completed within the test tube, achieving a single-tube operation; eliminating the step of transferring samples between multiple devices, resulting in a lower contamination rate and simpler operation compared to traditional methods, thereby improving the accuracy, convenience and reliability of detection. Attached Figure Description

[0024] Figure 1The image shown is a side view of a tubular bacteria / cell detection device in an embodiment, with the sealing cap in the position of closing the upper opening;

[0025] Figure 2 The image shown is a side view of a tubular bacteria / cell detection device in an embodiment, with the sealing cap in the open position.

[0026] Figure 3 The image shown is a cross-section of a tubular bacterial / cell detection device in the embodiment. Figure 1 ;

[0027] Figure 4 The image shown is a cross-section of a tubular bacterial / cell detection device in the embodiment. Figure 2 . Detailed Implementation

[0028] To further illustrate the various embodiments, the present invention provides accompanying drawings. These drawings are part of the disclosure of the present invention and are mainly used to illustrate the embodiments, and can be used in conjunction with the relevant descriptions in the specification to explain the operating principles of the embodiments. With reference to these drawings, those skilled in the art should be able to understand other possible implementations and the advantages of the present invention. Components in the drawings are not drawn to scale, and similar component symbols are generally used to represent similar components.

[0029] In the description of this utility model, terms such as "upper", "lower", "left", "right", "front", and "rear" are based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of description and simplification of operation, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this utility model.

[0030] The present invention will now be further described in conjunction with the accompanying drawings and specific embodiments.

[0031] Reference Figures 1 to 4As shown, this embodiment provides a single-tube bacterial / cell detection device, including a test tube body 10, a sealing cap 20, a filter membrane 40, and a three-way valve 30. The test tube body 10 has an upper opening 11, an upper chamber 12 connected to the upper opening 11, a transition chamber 13 located below the upper chamber 12 and connected to the upper chamber 12, and a reaction chamber 15 and a waste liquid chamber 14 located below the transition chamber 13 and connected to the transition chamber 13 respectively. Specifically, in this embodiment, the waste liquid chamber 14 and the reaction chamber 15 are distributed left and right. The sealing cap 20 is used to seal and cover the upper opening 11 of the test tube body 10. In this embodiment, the sealing cap 20 and the upper opening 11 of the test tube body 10 are connected by threads to form a sealing fit. That is, the sealing cap 20 has an internal thread 21, and the upper opening 11 of the test tube body 10 is provided with an external thread 111. The sealing cap 20 is screwed onto the external thread 111 of the test tube body 10 through the internal thread 21 to form a seal. The screw connection method simplifies the steps of closing and opening. Of course, in other embodiments, the sealing cap 20 can also seal the upper opening 11 by inserting it into the upper opening 11, etc.

[0032] The filter membrane 40 is disposed in the upper chamber 12. Specifically, in this embodiment, the filter membrane 40 is disposed at the lower outlet of the upper chamber 12 to trap bacteria or cells. The three-way valve 30 is disposed in the transition chamber 13 to control the connection and disconnection between the upper chamber 12, the reaction chamber 15 and the waste liquid chamber 14. In this embodiment, the three-way valve 30 can block the lower outlet of the upper chamber 12, connect the upper chamber 12 and the reaction chamber 15, and connect the upper chamber 12 and the waste liquid chamber 14.

[0033] Specifically, the test tube body 10 is made of a light-transmitting material or has a light-transmitting sidewall on the corresponding reaction chamber 15, for external detection instruments to detect it. In this embodiment, the test tube body 10 is made of transparent silicone, which is light-transmitting and not easily broken; its bottom has a high-transmittance area 151, which is part of the reaction chamber 15. Of course, in other embodiments, the test tube body 10 can also be made of a light-transmitting material such as glass, or it can be made of an opaque material, but the high-transmittance area 151 at the bottom is made of a light-transmitting material.

[0034] One preferred operating method for this single-tube bacterial / cell detection device is as follows:

[0035] Step 1: Filtration and collection of bacteria / cells: Unscrew the sealing cap 20 and inject the test solution into the upper chamber through the upper opening 11; control the three-way valve 30 to connect the upper chamber 12 and the waste liquid chamber 14, the test solution flows through the filter membrane 40, and the bacteria or cells in the test solution are trapped on the filter membrane 40; the waste liquid flows to the waste liquid chamber 14; after completion, control the three-way valve 30 to close the upper chamber 12;

[0036] Step 2, Lysis of bacteria / cells: Add lysis buffer to upper chamber 12 to infiltrate bacteria or cells; allow bacteria or cells to lyse and release ATP;

[0037] Step 3: Directing the flow to the reaction chamber: Control the three-way valve 30 to connect the upper chamber 12 and the reaction chamber 15, and the lysate carrying ATP flows into the reaction chamber 15;

[0038] Step 4, ATP fluorescence detection: Inject fluorescent detection reagent (such as luciferase or luciferin); the reagent reacts with ATP in the reaction chamber 15 to produce a light emission reaction; the external detection instrument reads the fluorescence intensity through the test tube (specifically through the high-transmittance zone 151 at the bottom) to calculate the concentration of bacteria or cells.

[0039] The entire process from collection and lysis of the bacteria / cells (i.e., bacteria or cells) to detection is completed within the test tube 10, achieving a single-tube operation; eliminating the step of transferring samples between multiple devices, resulting in a lower contamination rate and simpler operation compared to traditional methods, thereby improving the accuracy, convenience and reliability of detection.

[0040] Specifically, in this embodiment, the three-way valve 30 is a manual valve, specifically an L-shaped three-way ball valve. The three-way valve 30 is connected to a knob 31, which is externally mounted on the test tube body 10. Manual operation is performed via the external knob 31. For easier identification, the knob 31 is marked with symbols such as arrows to indicate the current position of the three-way valve 30. The external knob 31 needs to pass through the side wall of the test tube body 10, requiring a high degree of sealing in the assembly. Of course, in other embodiments, a pneumatic three-way valve 30 can also be used, that is, a three-way valve 30 extends from the side wall of the test tube body 10 through an air tube, and the three-way valve 30 is switched by an external pneumatic device; or the three-way valve 30 can be a remotely controllable electric valve, which has a built-in battery and control system, etc. The three-way valve 30 can be completely encapsulated in the test tube body 10, without the test tube body 10 being opened; the electric valve forms a wireless communication connection with the remote end, and the operation of the three-way valve 30 is controlled by the remote end. Specifically, the remotely controllable electric valve is an existing structure, which can use a micro motor to drive the ball valve to act, or it can be a three-way solenoid valve, etc., as long as it can control the above-mentioned chambers to switch and connect.

[0041] Specifically, in this embodiment, a fluorescent detection reagent can be pre-injected into the reaction chamber 15. The fluorescent detection reagent can be in liquid or lyophilized form. After the lysis buffer flows in, it automatically dissolves the reagent and triggers the reaction, eliminating the need for manual sample addition. This further reduces the number of operation steps and lowers the risk of contamination.

[0042] In this embodiment, the test tube body 10 is made of transparent silicone, which has a certain degree of flexibility. It can drive the sample to pass through the filter membrane 40 quickly by squeezing the tube wall to generate a pressure difference, making it suitable for rapid field detection.

[0043] Furthermore, the upper chamber 12 is also provided with a pre-filter membrane (not shown) with a pore size larger than that of the filter membrane; this pre-filter membrane is used to filter impurities. The lower filter membrane 40 is dedicated to the capture of bacteria or cells, improving the detection accuracy of complex samples (such as sewage, food homogenates).

[0044] The bottom of the test tube body 10 is also provided with a flat-bottomed upright part 16, and the high-transparency area 151 is located inside the upright part 16. The flat-bottomed upright part 16 allows the test tube body 10 to be placed upright, thereby effectively positioning the high-transparency area 151, so that the high-transparency area 151 of the reaction chamber 15 can directly correspond to the reading window of the fluorescence detector without the need for an auxiliary support.

[0045] In a further preferred embodiment, magnetic nanoparticles can be disposed on the surface of the filter membrane 40. An external magnetic field is applied during lysis to enhance the efficiency of bacterial or cell disruption. A magnet is also embedded at the bottom of the reaction chamber 15 to enrich the magnetic beads and avoid interference with optical detection.

[0046] Although the present invention has been specifically shown and described in conjunction with preferred embodiments, those skilled in the art should understand that various changes in form and detail may be made to the present invention without departing from the spirit and scope of the present invention as defined in the appended claims, and all such changes shall be within the scope of protection of the present invention.

Claims

1. A single-tube bacterial / cell detection device, characterized in that: The test tube includes a test tube body, a sealing cap, a filter membrane, and a three-way valve. The test tube body has an upper opening, an upper chamber connected to the upper opening, a transition chamber located below the upper chamber and connected to the upper chamber, and a reaction chamber and a waste liquid chamber located below the transition chamber and connected to the transition chamber respectively. The sealing cap is used to seal the upper opening of the test tube body; the filter membrane is disposed in the upper chamber; the three-way valve is disposed in the transition chamber to control the flow between the upper chamber, the reaction chamber and the waste liquid chamber.

2. The single-tube bacterial / cell detection device according to claim 1, characterized in that: The three-way valve is a manual valve, and the three-way valve is connected to a knob, which is externally located on the test tube body.

3. The single-tube bacterial / cell detection device according to claim 1, characterized in that: The three-way valve is a remotely controllable electric valve.

4. The single-tube bacterial / cell detection device according to claim 1, characterized in that: The three-way valve is an L-type three-way ball valve.

5. The single-tube bacterial / cell detection device according to claim 1, characterized in that: The test tube is made of transparent silicone.

6. The single-tube bacterial / cell detection device according to claim 1, characterized in that: The upper opening of the test tube body is connected to the sealing cap by a thread, forming a sealing fit.

7. The single-tube bacterial / cell detection device according to claim 1, characterized in that: The reaction chamber and the waste liquid chamber are distributed on the left and right sides.

8. The single-tube bacterial / cell detection device according to claim 1 or 7, characterized in that: The bottom of the test tube has a high-transmittance zone, which is part of the reaction chamber.

9. The single-tube bacterial / cell detection device according to claim 8, characterized in that: The bottom of the test tube is also provided with a flat-bottomed upright section, and the high-transparency zone is located inside the upright section.

10. The single-tube bacterial / cell detection device according to claim 1, characterized in that: The upper chamber is also equipped with a pre-filter membrane with a pore size larger than that of the filter membrane.

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