Membrane bacteria collection flaking method

Abstract

The invention discloses a film bacteria-collecting slide preparation method, which belongs to the field of medical detection, and comprises the following steps: step 1, preparing a specimen required by detection; 2, a sample is sucked through a concentration slide making device, the concentration slide making device comprises a membrane carrier and a bacterium collector which are used in a matched mode, the bacterium collector comprises a sampling pipe and a sampling part which are communicated in a sealed mode, a flow channel with an open end face is formed in the sampling part, the sampling part is matched with the membrane carrier, a microfiltration membrane is arranged at one end of the membrane carrier, and the microfiltration membrane is arranged at the other end of the membrane carrier; the microfiltration membrane is immersed in the specimen, the specimen enters the sampling tube under the action of negative pressure, and in the process, an object to be detected in the specimen is intercepted by the microfiltration membrane; and step 3, transferring the microfiltration membrane to the slide glass, and then separating the microfiltration membrane from the membrane carrier. The invention aims to reduce the cost of cell and bacterium detection.

Description

Technical Field

[0001] The invention relates to the field of medical detection, in particular to a method for preparing membrane bacteria slices. Background Art

[0002] When using the filter membrane (microporous filter membrane) recorded in patents such as announcement number CN102580548B for cell and bacteria detection, it is necessary to use the technical means recorded in patents such as announcement number CN106248978B to intercept bacteria and cells. Subsequently, the microporous filter membrane is fixed on a glass slide using the technical solution proposed in patents such as announcement number CN108760458B for subsequent operations such as microscopy. It can be seen that under the existing technical framework, the use of microporous filter membranes for cell and bacteria detection not only has high equipment costs, but also puts forward corresponding strict requirements on the detection site, resulting in a high detection cost of the microporous filter membrane, which to a certain extent limits the widespread promotion and application of microporous filter membrane technology. Summary of the invention

[0003] The purpose of the present invention is to provide a method for preparing membrane-collected bacteria slices in order to solve the above problems, so as to reduce the cost of cell and bacteria detection.

[0004] To achieve the above purpose, the technical solution adopted by the present invention is: a method for preparing membrane-collected bacteria slices, which comprises the following steps:

[0005] 1) Prepare specimens required for testing;

[0006] 2) The specimen is sucked by a concentration slide maker, the concentration slide maker includes a matching membrane carrier and a bacteria collector, the bacteria collector includes a sealed and connected sampling tube and a sampling part, the sampling part is provided with a flow channel with an open end face, the sampling part is adapted to the membrane carrier, one end of the membrane carrier is provided with a microporous filter membrane, the microporous filter membrane is immersed in the specimen, and the specimen enters the sampling tube under the action of negative pressure. In this process, the object to be detected in the specimen is intercepted by the microporous filter membrane;

[0007] 3) Transfer the microporous filter membrane to a slide, and then separate the microporous filter membrane from the membrane carrier.

[0008] Furthermore, one end of the sampling tube is open, and a rubber plug is provided at the open position. The internal pressure of the sampling tube is lower than the air pressure. The rubber plug is provided with a groove adapted to the sampling needle. The sampling needle is arranged on the sampling part and is connected with the flow channel. When the specimen is absorbed, it is only necessary to make the sampling needle pierce the rubber plug. Since the pressure of the outside air is higher than the pressure in the sampling cylinder, the specimen enters the sampling tube.

[0009] Furthermore, an air hole is provided at one end of the sampling tube, and the sampling tube is sealed and connected to the sampling part at one end of the air hole, and the air hole is connected to the flow channel; a vacuum pumping mechanism is provided in the sampling tube; and the specimen is sucked into the sampling tube through the vacuum pumping mechanism or the exhaust device.

[0010] Furthermore, in order to reduce the dependence on other supporting equipment and facilitate use in places with relatively simple conditions such as outdoors, a vacuum mechanism is provided in the sampling tube to extract the specimen, and the vacuum mechanism includes a piston adapted to the inner wall of the sampling tube, and the piston is connected to the push rod. The specimen is extracted by pulling the piston.

[0011] Furthermore, the sampling tube and the sampling part are separately arranged, and the sampling tube is provided with a connecting tube with an open end face on the outer side of the pore, and the connecting tube is nested and connected with the flow channel, so that the sampling tube and the sampling part are also produced separately, which reduces the difficulty of production and ensures that the required specimen can be intercepted by the microporous filter membrane.

[0012] Furthermore, in order to improve the level of automation, the sampling tube is connected to the air extraction device, and a connecting tube inserted into the flow channel is provided on the sampling tube.

[0013] Furthermore, an adhesive layer is provided on the carrier sheet, and the microporous filter membrane is fixed on the carrier sheet by gluing.

[0014] Furthermore, in order to fix the microporous filter membrane on the slide, a heating part is arranged at the lower end of the sampling part, a control button for controlling the operation of the heating part and the air extraction device is arranged on the gripping part, and the sampling tube is arranged in the gripping part. The microporous filter membrane is melted by the heating part, and then the microporous filter membrane is bonded to the slide.

[0015] Furthermore, in order to assemble the sampling tube and the holding part together and keep the soft rubber material away from the heating part, one end of the holding part is open, and a soft rubber part is provided at one end of the sampling part. The soft rubber part is adapted to the open end of the holding part, and the flow channel on the soft rubber part and the sampling part are connected.

[0016] Furthermore, in order to increase the sampling area, the sampling portion is provided with a countersunk hole at one end of the membrane carrier, and the opening of the flow channel at one end of the membrane carrier is located at the bottom of the countersunk hole. The sampling area is expanded through the countersunk hole. When using the microporous filter membrane described in patents such as CN102580548B, in order to avoid the rupture of the microporous filter membrane, a support portion in contact with the microporous filter membrane is provided at the lower end of the countersunk hole, and channels are provided on the support portion at intervals. When extracting the specimen, the support portion supports the microporous filter membrane to avoid the rupture of the microporous filter membrane due to excessive deformation.

[0017] Beneficial effects of the present invention: The present invention provides a membrane bacteria collection and preparation method. When the specimen is absorbed, the bacteria collector is assembled with the membrane carrier, and then the microporous filter membrane is immersed in the specimen. The inactivated specimen (liquid or gaseous) enters the sampling tube through the microporous filter membrane under negative pressure, and the cells and bacteria in the specimen are intercepted by the microporous filter membrane; then, the microporous filter membrane is pressed and fixed on a glass slide to quickly complete the preparation, thereby reducing the cost of using the microporous filter membrane for cell and bacteria detection. In this way, sampling and detection can be separated. When necessary, a large number of collection points can be quickly set up, and these collection points can complete the collection and preparation of bacteria through relatively simple operations. Then, the specimens will be collected and tested in a unified manner, so that a long-term investigation mechanism for the source of infection can be established, which not only improves the detection efficiency, but also reduces the detection cost. At the same time, centralized staining and reading of films also facilitates statistical filing work and ensures the quality of detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Schematic diagram of the membrane carrier structure.

[0019] Figure 2 This is a schematic diagram of the structure of the bacteria collector in Example 1.

[0020] Figure 3 This is a schematic diagram of the structure of the bacteria collector in Example 2.

[0021] Figure 4 This is a schematic diagram of the structure of the bacteria collector in Example 3.

[0022] Figure 5 This is a schematic diagram of the structure in the film-making state in Example 3.

[0023] The text labels in the figure are as follows: 1. membrane carrier; 2. sampling tube; 3. sampling part; 4. flow channel; 5. microporous filter membrane; 6. carrier; 7. rubber plug; 8. sampling needle; 9. piston; 10. push rod; 11. connecting tube; 12. holding part; 13. soft rubber part; 14. heating part; 15. control button; 16. supporting part; 17. connecting tube. DETAILED DESCRIPTION

[0024] In order to enable those skilled in the art to better understand the technical solution of the present invention, the present invention is described in detail below in conjunction with the accompanying drawings. The description in this part is only exemplary and explanatory and should not have any limiting effect on the protection scope of the present invention.

[0025] Embodiment 1, as Figure 1-2 As shown, the specific structure of the present invention is a method for preparing membrane bacteria slices, which comprises the following steps:

[0026] 1) Prepare the specimens required for the test. The specific specimen preparation process is carried out according to the existing specifications based on the items to be tested. In this embodiment, the detection of Mycobacterium tuberculosis is used as an example for explanation. The preparation process includes the subject spitting sputum into the specimen cup, and the tester adding 2 to 4 times the amount of digestion and inactivation liquid to the specimen, mixing it evenly for thirty minutes. The digestion and inactivation liquid contains sodium hypochlorite, etc.;

[0027] 2) The specimen is absorbed by a concentration slide maker, the concentration slide maker includes a matching membrane carrier 1 and a bacteria collector, the bacteria collector includes a sampling tube 2 and a sampling part 3, the sampling part 3 is provided with a flow channel 4 with an open end face, the sampling part 3 is adapted to the membrane carrier 1, in this embodiment, the membrane carrier 1 is a truncated cone structure with open ends, one end of the membrane carrier 1 is provided with a microporous filter membrane 5, the microporous filter membrane 5 is immersed in the specimen, and the specimen enters the sampling tube 2 under the action of negative pressure. In this process, the object to be detected in the specimen is retained by the microporous filter membrane 5; the microporous filter membrane 5 can adopt the filter membrane recorded in Patent No. CN102580548B.

[0028] The sampling part 3 is provided with a countersunk hole at one end of the membrane carrier 1, and the flow channel 4 is located at the bottom of the countersunk hole at the opening at one end of the membrane carrier 1. A support part 16 for supporting the microporous filter membrane 5 is provided at the lower end of the countersunk hole, and channels are provided on the support part 16 at intervals.

[0029] 3) The microporous filter membrane 5 is transferred to the slide 6, and then the microporous filter membrane 5 is separated from the membrane carrier 1. The slide 6 carrying the microporous filter membrane 5 is then stored in a specimen box. The test personnel collect the slides 6 to perform operations such as staining and reading the microporous filter membrane 5. The slide 6 can be a glass slide or the like.

[0030] In this embodiment, one end of the sampling tube 2 is open, and a rubber plug 7 is provided at the open position. The internal pressure of the sampling tube 2 is less than the air pressure. The rubber plug 7 is provided with a groove adapted to the sampling needle 8. In order to reduce the procurement cost, the sampling tube 2 can directly use a vacuum blood collection tube that meets the requirements. The sampling needle 8 is provided on the sampling part 3 and communicates with the flow channel 4. Under normal conditions, the sampling needle 8 does not pierce the rubber plug 7, and a certain distance is provided between the rubber plug 7 and the sampling part 3. The sampling part 3 is made of a soft rubber material or the outside of the sampling part 3 is covered with a soft rubber layer. The soft rubber material and the soft rubber layer can be made of silicone, rubber, etc. A circle of baffles concentric with it can be provided at the upper end of the sampling part 3, and the lower end of the sampling tube 2 is inserted into the space enclosed by the baffle, and the outer diameter of the lower end of the sampling tube 2 is adapted to the inner diameter of the baffle. An adhesive layer is provided on the slide 6, and the adhesive layer is made of glue, double-sided tape, etc. that meet the hygiene requirements.

[0031] Specific method of use: After collecting and inactivating the specimen, install the membrane carrier 1 on the sampling part 3. After the sampling part 3 and the membrane carrier 1 are assembled, the lower end of the support part 16 will support the upper end of the microporous filter membrane 5. Then immerse the microporous filter membrane 5 in the specimen, and push the sampling tube 2 downward so that the sampling needle 8 pierces the rubber plug 7. At this time, the sampling tube 2 is connected to the outside world. Under the action of negative pressure, the specimen is extracted into the sampling tube 2, and the cells and bacteria in the specimen are intercepted on the microporous filter membrane 5, thereby completing the absorption of the specimen. When making the slice, press the microporous filter membrane 5 on the adhesive layer, so that the microporous filter membrane 5 is fixed on the slide 6, and then lift the membrane carrier 1 upward to separate the microporous filter membrane 5 and the membrane carrier 1, thereby completing the slice making.

[0032] Embodiment 2, as Figure 1 , Figure 3 As shown, the other steps and structures of this embodiment are the same as those of embodiment 1, but the sampling tube in this embodiment is different from that in embodiment 1. In this embodiment, an air hole is provided at one end of the sampling tube 2, and the sampling tube 2 is sealed and connected to the sampling part 3 at one end of the air hole, and the air hole is connected to the flow channel 4; a vacuum mechanism is provided in the sampling tube 2. The vacuum mechanism includes a piston 9 adapted to the inner wall of the sampling tube 2, and the piston 9 is connected to a push rod 10 at the far end of the air hole, and the push rod 10 extends out of the sampling tube 2, and the piston 9 is tightly fitted to the inner wall of the sampling tube 2. The specimen is extracted by moving the piston 9, and a sealing ring is provided on the edge of the piston 9 to ensure the sealing during the specimen extraction process.

[0033] The sampling part 3 is made of a soft rubber material such as silicone or the outer wall of the sampling part 3 is covered with a soft rubber layer made of a soft rubber material such as silicone or rubber. The outer diameter of the sampling part 3 and the inner diameter of the membrane carrier 1 can be interference fit. Since the sampling part 3 and the sampling tube 2 are made of two different materials, the sampling tube 2 and the sampling part 3 are separately provided for the convenience of production. A connecting tube 11 is provided on the outer side of the pore of the sampling tube 2. When assembling, the connecting tube 11 is inserted into the flow channel, and the end face of the connecting tube 11 is open. An adhesive layer is provided on the carrier 6.

[0034] Specific method of use: After collecting and inactivating the specimen, install the membrane carrier 1 on the sampling part 3. After the sampling part 3 and the membrane carrier 1 are assembled, the lower end of the support part 16 will support the upper end of the microporous filter membrane 5. Then immerse the microporous filter membrane 5 in the specimen, and pull the piston 9 upward through the push rod 10. The negative pressure generated will extract the specimen into the sampling tube 2. At this time, the cells and bacteria in the specimen are intercepted on the microporous filter membrane 5, thereby completing the absorption of the specimen. When making a slice, press the microporous filter membrane 5 on the adhesive layer, so that the microporous filter membrane 5 is fixed on the slide 6, and then lift the membrane carrier 1 upward to separate the microporous filter membrane 5 and the membrane carrier 1, thereby completing the slice making.

[0035] Embodiment 3, as Figure 1 , Figure 4 , Figure 5 As shown, the other steps and structures of this embodiment are the same as those of embodiment 1, but in this embodiment, the upper end of the sampling tube 2 is connected to the air extraction device (not shown in the figure) through a pipeline, and the air extraction device can adopt existing equipment such as existing vacuum pumps and fans that can extract air and blow out airflow, or the air extraction device includes an exhaust pipe sealed with the pipeline, and a working piston adapted thereto is arranged in the exhaust pipe, and the working piston moves along the exhaust axis under the action of existing linear drive devices such as electric push rods and linear modules. When the working piston is away from one end of the pipeline, it is exhausting, and when the piston is close to the pipeline, air is blown into the sampling tube 2.

[0036] The sampling tube 2 is arranged in the gripping part 12, one end of the gripping part 12 is open, and one end of the sampling part 3 is provided with a soft rubber part 13, which can be made of soft rubber materials such as silicone, rubber, thermoplastic polyurethane elastomer, etc. The outer diameter of the soft rubber part 13 is interference fit with the inner diameter of the open end of the gripping part 12, and the flow channel 4 on the soft rubber part 13 is connected with the flow channel 4 on the sampling part 3. The lower end of the sampling tube 2 is provided with a connecting tube 17, which is inserted into the flow channel 4, and the flow channel 4 and the outer wall of the connecting tube 17 are interference fit. The soft rubber part 13 is provided to ensure the connection between the sampling part 3 and the gripping part 12, and the soft rubber part 13 is away from the heating part 14.

[0037] The heating part 14 is arranged in a ring shape at the lower end of the sampling part 3. When the heating part 14 is working, the microporous filter membrane 5 can be melted. The heating part 14 can adopt an existing heating structure such as a heating wire. This embodiment takes the use of a heating wire as an example. The heating wire is connected to an electric wire. A channel for the wire to pass through is provided in the sampling part 3 and the soft rubber part 13. The wire passes through the sampling part 3 and the soft rubber part 13 in turn and extends into the holding part 12 and is connected to the control circuit board in the holding part 12. The power interface of the control circuit board is connected to the plug through the wire. A control button 15 for controlling the operation of the heating part 14 and the air extraction device is provided on the holding part 12 and the control button 15 is connected to the control input end of the control circuit board. A plurality of control buttons 15 are provided to control the heating part 14 and the air extraction device respectively; electronic components such as a control chip are provided on the control circuit board.

[0038] Specific method of use: After collecting and inactivating the specimen, install the membrane carrier 1 on the sampling part 3, and then immerse the microporous filter membrane 5 in the specimen. Through the negative pressure generated by the air extraction device, the specimen passes through the microporous filter membrane 5, the flow channel 4, and the connecting tube 17 in sequence into the sampling tube 2, and the cells, bacteria, etc. in the specimen are intercepted on the microporous filter membrane 5. After the specimen is absorbed, the tester places the microporous filter membrane 5 on the slide 6, and then presses the heating part 14 flatly on the upper end of the microporous filter membrane 5. With the operation of the heating part 14 and the pressure applied by the tester to the microporous filter membrane 5, the microporous filter membrane 5 is fixed on the slide 6, and then the membrane carrier 1 is lifted upward to separate the microporous filter membrane 5 and the membrane carrier 1, thereby completing the preparation.

[0039] Embodiment 4, as Figure 1 , Figure 4 , Figure 5 As shown, the other steps and structures of this embodiment are the same as those of embodiment 3, but in this embodiment, the lower end of the sampling portion 3 is not provided with a heating portion 14, but an adhesive layer is provided on the supporting slide 6. When making the slide, the microporous filter membrane 5 is pressed on the adhesive layer, so that the microporous filter membrane 5 is fixed on the slide 6 such as a slide, thereby completing the slide making.

[0040] It should be noted that, in this article, the terms "comprises", "includes" or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article or apparatus that includes a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or apparatus.

[0041] This article uses specific examples to illustrate the principles and implementation methods of the present invention. The above examples are only used to help understand the method and core ideas of the present invention. The above is only a preferred implementation of the present invention. It should be pointed out that due to the limitations of textual expression and the objective existence of infinite specific structures, ordinary technicians in this technical field can make several improvements, modifications or changes without departing from the principles of the present invention, and can also combine the above technical features in an appropriate manner; these improvements, modifications, changes or combinations, or the direct application of the inventive concept and technical solution to other occasions without improvement, should be regarded as the protection scope of the present invention.

Claims

1. A method for preparing membrane-collected bacteria slices, characterized in that: The following steps are involved: 1) Prepare specimens required for testing; 2) absorbing the specimen through a concentration slide maker, the concentration slide maker comprising a matching membrane carrier (1) and a bacteria collector, the bacteria collector comprising a sealed and connected sampling tube (2) and a sampling portion (3), the sampling portion (3) being provided with a flow channel (4) with an open end face, the sampling portion (3) being adapted to the membrane carrier (1), one end of the membrane carrier (1) being provided with a microporous filter membrane (5), the microporous filter membrane (5) being immersed in the specimen, the specimen entering the sampling tube (2) under the action of negative pressure, during which the object to be detected in the specimen is retained by the microporous filter membrane (5); 3) The microporous filter membrane (5) is transferred to a carrier sheet (6), and then the microporous filter membrane (5) is separated from the membrane carrier (1).

2. A method for preparing membrane-collected bacteria slices according to claim 1, characterized in that: One end of the sampling tube (2) is open, and a rubber plug (7) is provided at the open end. The internal pressure of the sampling tube (2) is lower than the air pressure. The rubber plug (7) is provided with a groove adapted to a sampling needle (8). The sampling needle (8) is provided on the sampling portion (3) and is connected to the flow channel (4).

3. The method for preparing membrane-collected bacteria slices according to claim 1, characterized in that: An air hole is provided at one end of the sampling tube (2); the sampling tube (2) is sealedly connected to the sampling portion (3) at one end of the air hole; the air hole is communicated with the flow channel (4); and a vacuum extraction mechanism is provided inside the sampling tube (2).

4. A method for preparing membrane-collected bacteria slices according to claim 3, characterized in that: The vacuum extraction mechanism comprises a piston (9) adapted to the inner wall of the sampling tube (2), wherein the piston (9) is connected to a push rod (10) at the far end of the air hole, and the push rod (10) extends out of the sampling tube (2).

5. The method for preparing membrane-collected bacteria slices according to claim 3, characterized in that: The sampling tube (2) and the sampling portion (3) are arranged separately, and the sampling tube (2) is provided with a connecting tube (11) with an open end surface at the outer side of the air hole, and the connecting tube (11) is nested and connected with the flow channel (4).

6. The method for preparing membrane-collected bacteria slices according to claim 1, characterized in that: The sampling tube (2) is connected to the air extraction device, and a connecting tube (17) inserted into the flow channel (4) is provided on the sampling tube (2).

7. A method for preparing membrane-collected bacteria slices according to any one of claims 1 to 6, characterized in that: An adhesive layer is provided on the carrier sheet (6).

8. The method for preparing membrane-collected bacteria slices according to claim 6, characterized in that: A heating portion (14) is arranged at the lower end of the sampling portion (3), a control button (15) for controlling the operation of the heating portion (14) and the air extraction device is arranged on the holding portion (12), and the sampling tube (2) is arranged in the holding portion (12).

9. A method for preparing membrane-collected bacteria slices according to claim 8, characterized in that: One end of the holding portion (12) is open, and one end of the sampling portion (3) is provided with a soft rubber portion (13), the soft rubber portion (13) is adapted to the open end of the holding portion (12), and the soft rubber portion (13) is connected to a flow channel on the sampling portion (3).

10. The method for preparing membrane-collected bacteria slices according to claim 1, characterized in that: The sampling portion (3) is provided with a countersunk hole at one end of the membrane carrier, the flow channel (4) is located at the bottom of the countersunk hole at an open portion at one end of the membrane carrier (1), a support portion (16) in contact with the microporous filter membrane (5) is provided at the lower end of the countersunk hole, and channels are provided at intervals on the support portion (16).

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