Peristaltic pressure-controlled PRV (porcine reproductive virus) concentration system

Abstract

The invention discloses a peristaltic pressure control PRV virus concentration system which comprises a peristaltic pump, a virus collection bottle and a hollow fiber column which are sequentially connected in a closed loop mode, the two ends of a first rubber pipe on the peristaltic pump are connected with the virus collection bottle and the hollow fiber column respectively, and the hollow fiber column is connected with the virus collection bottle through a second rubber pipe. And one side of the hollow fiber column is connected with a waste liquid bottle through a third rubber pipe. The peristaltic pressure control PRV virus concentration system is simple to operate, high in circulating concentration efficiency, short in concentration time and low in personnel demand.

Description

Technical Field

[0001] The invention relates to the technical field of virus concentration, and in particular to a peristaltic pressure-controlled PRV virus concentration system. Background Art

[0002] Pseudorabies virus, also known as PRV, is an acute infectious disease of pigs that can cause abortion in pregnant sows, stillbirth, infertility in boars, death of newborn piglets, breathing difficulties in fattening pigs, growth stagnation and other symptoms. It is one of the major infectious diseases that endanger the global pig industry. In the study of PRV, it is first necessary to obtain a high concentration of the virus.

[0003] The existing PRV virus concentration method is mainly carried out by chemical flocculation. In the operation, it is necessary to discard the supernatant after ultracentrifugation, and then add a small amount of dissolving solution to achieve the purpose of concentration by reducing the volume of the dissolving solution. In this process, the centrifugal speed of the ultracentrifugation can reach 13000rpm / min, and the time is at least 30min, which has high requirements for instruments and equipment. In addition, the operation needs to be left to stand overnight to allow the PRV virus to flocculate, and then the operation is repeated, which takes a long time.

[0004] In addition, during the centrifugation process, the maximum size of the centrifuge tube used is generally 50mL / tube, and a batch can only hold 4-6 tubes, which makes it difficult to operate in large quantities and results in low work efficiency. Polyethylene glycol 8000 in the chemical flocculant needs to be dissolved before use, which not only takes a long time to dissolve, but also takes at least 22 hours to wait after adding the virus suspension, reducing work efficiency. Summary of the invention

[0005] The purpose of the present invention is to provide a peristaltic pressure-controlled PRV virus concentration system, which is simple to operate, has high circulation concentration efficiency, short concentration time and less personnel requirements.

[0006] To achieve the above-mentioned purpose, the present invention provides a peristaltic pressure-controlled PRV virus concentration system, comprising a peristaltic pump, a virus collection bottle and a hollow fiber column which are connected in a closed loop in sequence, wherein the two ends of a first rubber tube on the peristaltic pump are respectively connected to the virus collection bottle and the hollow fiber column, the hollow fiber column and the virus collection bottle are connected via a second rubber tube, and one side of the hollow fiber column is connected to a waste liquid bottle via a third rubber tube.

[0007] Preferably, the interior of the hollow fiber column is composed of hollow fiber filaments with openings at both ends, and both ends of the outer side are provided with outflow ports, and the outflow ports are connected to the waste liquid bottle through a third rubber tube.

[0008] Preferably, a liquid pressure control valve is provided on the second rubber tube.

[0009] Preferably, the level of the end of the second rubber tube in the virus collection bottle is higher than the level of the end of the first rubber tube.

[0010] Preferably, the liquid inlet at one end of the hollow fiber column is connected to a peristaltic pump, and the liquid outlet at the other end is connected to a liquid pressure control valve.

[0011] Preferably, the waste liquid bottle is close to the liquid outlet side of the hollow fiber column.

[0012] Therefore, the present invention adopts the above-mentioned peristaltic pressure-controlled PRV virus concentration system, which is simple to operate, has high circulation concentration efficiency, short concentration time, and less personnel requirements.

[0013] The technical solution of the present invention is further described in detail below through the accompanying drawings and embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is a structural schematic diagram of an embodiment of a peristaltic pressure-controlled PRV virus concentration system of the present invention.

[0015] Reference numerals

[0016] 1. First rubber tube; 2. Peristaltic pump; 3. Hollow fiber column; 4. Liquid inlet; 5. Liquid outlet; 6. Outflow port; 7. Second rubber tube; 8. Liquid pressure control valve; 9. Third rubber tube; 10. Virus collection bottle; 11. Waste liquid bottle. DETAILED DESCRIPTION

[0017] The technical solution of the present invention is further described below through the accompanying drawings and embodiments.

[0018] Unless otherwise defined, the technical terms or scientific terms used in the present invention should be understood by people with ordinary skills in the field to which the present invention belongs. The words "first", "second" and similar words used in the present invention do not indicate any order, quantity or importance, but are only used to distinguish different components. "Include" or "comprise" and similar words mean that the elements or objects appearing before the word include the elements or objects listed after the word and their equivalents, without excluding other elements or objects. "Connect" or "connected" and similar words are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect. "Up", "down", "left", "right" and the like are only used to indicate relative positional relationships. When the absolute position of the described object changes, the relative positional relationship may also change accordingly.

[0019] As shown in the figure, a peristaltic pressure-controlled PRV virus concentration system includes a peristaltic pump 2, a virus collection bottle 10 and a hollow fiber column 3 connected in a closed loop. The closed-loop connection design can achieve cyclic concentration, purify the virus hundreds of times, and the damage to the virus can be ignored, thereby improving work efficiency. The peristaltic pump 2 is used to load the virus liquid, which is simple to operate compared with complex equipment and does not require complex processing.

[0020] The two ends of the first rubber tube 1 on the peristaltic pump 2 are respectively connected to the virus collection bottle 10 and the hollow fiber column 3, the hollow fiber column 3 is connected to the virus collection bottle 10 through the second rubber tube 7, and one side of the hollow fiber column 3 is connected to the waste liquid bottle 11 through the third rubber tube 9. The interior of the hollow fiber column 3 is composed of hollow fiber filaments with openings at both ends, and both ends of the outer side are provided with outflow ports 6, and the outflow ports 6 are connected to the waste liquid bottle 11 through the third rubber tube 9. The liquid inlet 4 at one end of the hollow fiber column 3 is connected to the peristaltic pump 2, and the liquid outlet 5 at the other end is connected to the liquid pressure control valve 8. The PRV virus is a membrane virus with a size of about 180nm, which can be effectively blocked in the hollow fiber column 3 by the hollow fiber of 100kDa specification, and will not flow out from the outflow port 6 on the side of the hollow fiber column 3. The outflow port 6 on the outer side of the hollow fiber column 3 is used for the outflow of water and small molecular impurities in the virus liquid.

[0021] One end of the first rubber tube 1 is placed in the virus collection bottle 10, and the other end is connected to the liquid inlet 4 of the hollow fiber column 33. The first rubber tube 1 is stuck in the middle of the pump head of the peristaltic pump 2. The first rubber tube 1 filled with virus liquid is squeezed by the roller of the pump head of the peristaltic pump 2, and the virus liquid moves forward along the tube. The virus liquid is pumped by alternately squeezing and releasing the first rubber tube 1 of the elastic delivery hose of the pump. One end of the second rubber tube 7 is connected to the liquid outlet 5 of the hollow fiber column 3, and the other end is placed in the virus collection bottle 10. The virus liquid flowing out of the hollow fiber column 3 enters the virus collection bottle 10 through the second rubber tube 7. One end of the third rubber tube 9 is connected to the outflow port 6 near the liquid outlet 5 of the hollow fiber column 3, and the other end is placed in the waste liquid bottle 11. The third rubber tube 9 communicates the liquid outlet 5 and the waste liquid bottle 11.

[0022] The second rubber tube 7 is provided with a liquid pressure control valve 8. The liquid pressure control valve 8 is placed behind the liquid outlet 5 of the hollow fiber column 3. The liquid pressure valve generates pressure by squeezing the pipeline, forcing the water and small molecules in the virus liquid to be squeezed out from the outlet 6 on the side of the hollow fiber column 3. The water and small molecules of impurities flowing out of the outlet 6 enter the waste liquid bottle 11 through the third rubber tube 9. The waste liquid bottle 11 is close to the liquid outlet 5 of the hollow fiber column 3, which is convenient for collecting waste liquid.

[0023] The horizontal height of the end of the second rubber tube 7 in the virus collection bottle 10 is higher than the horizontal height of the end of the first rubber tube 1. The end of the first rubber tube 1 is closer to the bottom of the virus collection bottle 10 than the end of the second rubber tube 7. The end of the first rubber tube 1 first draws the virus liquid at the bottom of the virus collection bottle 10 to the peristaltic pump 2, and then the end of the second rubber tube 7 transports the virus liquid back to the virus collection bottle 10. This design is more conducive to the flow of the virus liquid in the rubber tube, and realizes circulation concentration.

[0024] The present invention adopts a peristaltic pressure-controlled PRV virus concentration system to concentrate viruses. The virus liquid is transported to a peristaltic pump 2 by a first rubber tube 1. The virus liquid moves forward along the tube to a liquid inlet 4 of a hollow fiber column 3 through the action of a pump head roller of the peristaltic pump 2. The hollow fiber column 3 effectively blocks the virus. Water and small molecular impurities are discharged into a waste liquid bottle 11 through an outflow port 6 of the hollow fiber column 3 through a third rubber tube 9. The remaining virus liquid is circulated back to a virus collection bottle 10 through a liquid outlet 5 of the hollow fiber column 3 through a second rubber tube 7. The virus liquid is recovered and then circulated and concentrated.

[0025] When the concentration system of the present invention is operated, the PRV virus of 1L is concentrated to 10 milliliters, and the titer (TCID50) of the virus increases by 2logs. In the whole process, only 20 minutes before the start and 1 minute after the virus concentrate is collected are operated, and the rest of the time is automatically carried out without human intervention. The total time has been used for about 4 hours, and the running time is short compared with the traditional treatment method. The virus collection bottle 10 is placed in a safety cabinet, and the laboratory and personnel are required to be small without the need for the experimenter to look after it at all times. The hollow fiber tangential flow mode is adopted to filter the virus, and the tangential flow refers to the filtering form in which the liquid flow direction and the filtering direction are vertical, and the force is different from the traditional terminal pressure filtration, and the pressure filter type is compared to concentrate, and the method for filtering the virus by tangential flow is not easy to block the hollow fiber silk, and small molecules can also be effectively filtered, and deionized water can be added to carry out purification treatment while concentrating. The concentration is carried out in the form of rear-end pressure control. The liquid pressure control valve 8 squeezes the pipeline, forcing water and small molecular impurities to flow out of the hollow fiber column 3 and be collected in the waste liquid bottle 11. The remaining virus liquid enters the virus collection bottle 10 and can be circulated and concentrated. It is not only efficient, but also does not require human supervision, complex instruments and equipment, and complex operations. It is simple and easy.

[0026] Therefore, the present invention adopts the above-mentioned peristaltic pressure-controlled PRV virus concentration system, which is simple to operate, has high circulation concentration efficiency, short concentration time, and less personnel requirements.

[0027] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that they can still modify or replace the technical solution of the present invention with equivalents, and these modifications or equivalent replacements cannot cause the modified technical solution to deviate from the spirit and scope of the technical solution of the present invention.

Claims

1. A peristaltic pressure-controlled PRV virus concentration system, characterized in that: The invention comprises a peristaltic pump, a virus collection bottle and a hollow fiber column which are connected in a closed loop in sequence. The two ends of a first rubber tube on the peristaltic pump are respectively connected to the virus collection bottle and the hollow fiber column. The hollow fiber column and the virus collection bottle are connected via a second rubber tube. One side of the hollow fiber column is connected to a waste liquid bottle via a third rubber tube.

2. A peristaltic pressure-controlled PRV virus concentration system according to claim 1, characterized in that: The interior of the hollow fiber column is composed of hollow fiber filaments with openings at both ends, and both ends of the outer side are provided with outflow ports, and the outflow ports are connected to the waste liquid bottle through a third rubber tube.

3. A peristaltic pressure-controlled PRV virus concentration system according to claim 1, characterized in that: The second rubber tube is provided with a liquid pressure control valve.

4. A peristaltic pressure-controlled PRV virus concentration system according to claim 1, characterized in that: The level of the end of the second rubber tube in the virus collection bottle is higher than the level of the end of the first rubber tube.

5. A peristaltic pressure-controlled PRV virus concentration system according to claim 3, characterized in that: The liquid inlet at one end of the hollow fiber column is connected to the peristaltic pump, and the liquid outlet at the other end is connected to the liquid pressure control valve.

6. A peristaltic pressure-controlled PRV virus concentration system according to claim 1, characterized in that: The waste liquid bottle is close to the liquid outlet side of the hollow fiber column.

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