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Hog fever cell-adapted virus, hog fever virus live vaccine prepared therefrom and preparation method thereof

A swine fever virus and live vaccine technology, applied in the field of swine fever virus live vaccine and preparation, can solve the problems of low virus content, high vaccine cost, less harvest times, etc., and achieve high virus content, large virus liquid volume, and immune efficacy. high effect

Active Publication Date: 2016-08-24
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method adopts ST cell line when propagating classical swine fever virus "C strain" to produce virus seed, but still needs to use a large amount of rabbits when propagating basic virus seed, and this method collects poison when the propagation of swine fever seedling venom The time is longer, the number of harvests is less, and the virus fluid harvested per unit cell is less
In addition, the method has low virus content and high vaccine cost

Method used

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  • Hog fever cell-adapted virus, hog fever virus live vaccine prepared therefrom and preparation method thereof
  • Hog fever cell-adapted virus, hog fever virus live vaccine prepared therefrom and preparation method thereof
  • Hog fever cell-adapted virus, hog fever virus live vaccine prepared therefrom and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Screening of hog fever rabbitized attenuated virus-adapted cells

[0040] 1. Viruses and cell lines

[0041] The attenuated swine fever rabbit virus is the P480 generation spleen virus (China Veterinary Drug Control Institute), and BT cells (China Veterinary Drug Control Institute) are used as cell lines for adapting the virus. ST cells (ATCC, American type culture collection, American type culture collection bank), PK15 cells (China Veterinary Drug Administration), IBRS-2 cells (China Veterinary Drug Administration), bovine testicular primary cells (self-made), SK6 cells (ATCC).

[0042] 2. Experimental method

[0043] (1) Proliferation test in cells of attenuated swine fever rabbit virus

[0044] ①Preparation method of bovine testicular cells

[0045] Aseptically take 1 pair of testicles from newborn calves, soak them in Hank's solution containing 1000U / ml penicillin, remove cysts and testicular capsule membranes under aseptic conditions, and then add 0...

Embodiment 2

[0057] Embodiment 2: Hog fever rabbitized attenuated cells adapt to the process of domestication

[0058] 1. Experimental materials

[0059] (1) Viruses and cell lines

[0060] The attenuated swine fever rabbit virus is the P480 generation spleen virus (China Veterinary Drug Control Institute), and BT cells (China Veterinary Drug Control Institute) are used as cell lines for adapting the virus.

[0061] (2) Test pigs

[0062] The pigs used in the experiment were all from rural free-range households, healthy and sensitive pigs that had no contact with CSFV and CSF vaccine. Blood was drawn from pigs one by one before use, and the neutralization antibody of swine fever was negative in the neutralization test. The detection of classical swine fever antigen, porcine circovirus type 2 and porcine reproductive disorder and respiratory syndrome virus by PCR method were all negative.

[0063] 2. Preparation method

[0064] (1) Preparation of BT monolayer cells

[0065] After the ...

Embodiment 3

[0080] Example 3: Determination of the response of classical swine fever virus cells to different generations of CC strains to induce body temperature in rabbits

[0081] 1. Test material

[0082] (1) Virus: CSF cell-adapted virus (P20 generation, P30 generation, P40 generation), swine fever rabbit attenuated virus (P480, China Veterinary Drug Control Institute);

[0083] (2) cells: bovine turbinate bone cells (BT cells);

[0084] (3) Experimental animals: 10 big-eared white rabbits aged 3-4 months (Henan Province Experimental Animal Center).

[0085] 2. Test method

[0086] (1) Determination of antibodies against classical swine fever in experimental animals

[0087] Indirect hemagglutination inhibition kit (Lanzhou Veterinary Research Institute) was used to measure the antibody titer of swine fever in rabbit serum, and the titer of swine fever antibody was less than 1:2 for the test animals.

[0088] (2) Indirect immunofluorescence assay method

[0089] BT cells in the ...

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Abstract

The invention relates to a swine fever cell-adapted virus, a live swine fever virus vaccine prepared therefrom and a preparation method thereof. In the method, the adaptive virus of classical swine fever virus is used as seedling virus seed, and the original virus seed, basic virus seed and production virus seed of classical classical virus are prepared by successive cell culture, and then the virus seed is produced by the adaptive virus of classical swine fever virus in the passaged cell. After multiplication, the qualified virus liquid is continuously harvested as the antigen for the attenuated swine fever vaccine, and the swine fever cell-adapted live vaccine is obtained through seedling preparation, subpackaging, and freeze-drying. The preparation of poison seeds solves animal welfare issues such as slaughtering rabbits for the prevention of swine fever, and reduces the possible risk of exogenous factors and potential shedding of poison in the preparation of spleen poison. The method for preparing a live vaccine of swine fever virus has the advantages of short virus propagation time, many harvest times, large amount of virus liquid harvested per unit cell, high virus content and low production cost. The prepared swine fever virus live vaccine has good safety and high immune efficacy.

Description

technical field [0001] The invention belongs to the technical field of animal biopharmaceuticals, and relates to a swine fever cell-adapted virus, a live swine fever virus vaccine prepared therefrom and a preparation method. Background technique [0002] Classical swine fever virus (CSFV) is a highly contagious disease caused by classical swine fever virus (CSFV). Pigs of different ages, sexes and breeds can be infected, and the mortality rate is as high as 80% to 90%. Swine fever is classified as one of the Class A diseases by the International Organization for Animal Health, and my country classifies it as a Class I disease. The attenuated swine fever rabbit vaccine successfully developed by Chinese scholars is currently recognized as a safe and effective vaccine in the world. With the help of intensive vaccination of C-strain vaccine and comprehensive prevention and control measures, relevant countries have effectively controlled swine fever and even eliminated swine feve...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12N7/08A61K39/187A61P31/14C12R1/93
Inventor 张许科孙进忠白朝勇
Owner PU LIKE BIO ENG
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