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Method for producing swine fever vaccines

A live swine fever vaccine and production method technology, applied in the field of veterinary biological products, can solve the problems of exogenous virus contamination risk, high quality control difficulty, poor genetic stability of virus, etc., to improve genetic stability and biological safety , high content of active ingredients in the vaccine, stable immunogenicity of the vaccine

Inactive Publication Date: 2012-09-05
CHINA INST OF VETERINARY DRUG CONTROL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, GPE strain and Thiverval strain are produced by pig-derived cell lines, which have a greater risk of known or unknown exogenous virus contamination. Once they are not well controlled, they can easily lead to certain existing or newly discovered infectious diseases popular
Among them, there are currently two production processes for rabbitized attenuated live vaccines. The first process is to use rabbits to produce vaccines. The tissue or lymph nodes or spleens of rabbits inoculated are used to make vaccines. This process can effectively avoid foreign virus contamination and ensure the virus The genetic stability of the virus does not become stronger, but it needs to use a large number of rabbits, the quality control is difficult and the production cost is high; the second process is the production of cattle, sheep primary cells or pig passage cells. Avoid using a large number of experimental animals, but there is a risk of exogenous virus contamination (because bovine, sheep primary cells or pig passage cells are susceptible to many known or unknown exogenous viruses) and the genetic stability of the virus is slightly poor, and the virus has an independent The risk of returning to strength
In addition, the attenuated swine fever rabbit virus is domesticated through continuous passage of rabbits, and the seed virus has not been cloned and purified. Therefore, different batches of seed poisons are different from the same batch of rabbits or the same batch of seed poisons from different batches. The infectivity and immunogenicity of rabbits often have certain differences

Method used

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  • Method for producing swine fever vaccines
  • Method for producing swine fever vaccines
  • Method for producing swine fever vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Cloning and Identification of Example 1 Labinized Attenuated Virus of Classical Swine Fever

[0056] 1. Cloning of the attenuated rabbit virus of classical swine fever

[0057] Carried out with viral gene rescue.

[0058] (1) Viral RNA extraction: TRIzol Reagent kit (purchased from Invitrogen Company) was used to extract the total RNA of hog fever rabbitized attenuated cell culture medium;

[0059] (2) Whole genome cDNA clone:

[0060] 1) Design 7 pairs of primers (sequences 1-14, Table 1) to carry out RT-PCR amplification respectively, and the amplified products are purified and recovered respectively, and the amplified products are detected by agarose electrophoresis ( figure 1 ), the target band appeared as a result, indicating that the amplification was successful;

[0061] Table 1 is used to amplify the primer sequence of the full genome of classical swine fever virus rabbitization attenuated strain (cell source)

[0062]

[0063] 2) Cloning the purified an...

Embodiment 2

[0080] The preparation of embodiment 2 vaccine

[0081] 1. Reproduction of seed poison

[0082] The well-grown RK-13 ​​seed batch cells were inoculated with CSFV CC-2 seed batch seed virus, and the virus was harvested after 3 to 4 days of cultivation, and used as the seed virus for production;

[0083] Seed poison test: test according to the provisions of "Chinese Veterinary Pharmacopoeia", and the results meet the requirements of sterility test and foreign virus test standards;

[0084] 2. Subculture and culture of production cells

[0085] After the seed batch cells RK-13 ​​are digested and dispersed, the cells are cultured in suspension, and when they grow to a suitable density, they are used for further passage or virus inoculation;

[0086] Cell test: test according to the provisions of "Chinese Veterinary Pharmacopoeia", and the results meet the requirements of cell standards for production and inspection;

[0087] 3. Propagation of virus fluid for production

[0088...

Embodiment 3

[0093] The preparation of embodiment 3 vaccines

[0094] 1. Reproduction of seed poison

[0095] Inoculate well-grown SK-6 seed batch cells with the CSFV CC-2 seed batch seed virus, and collect the virus after 3 to 4 days of cultivation, as the seed virus for production;

[0096] Seed poison test: test according to the provisions of "Chinese Veterinary Pharmacopoeia", and the results meet the requirements of sterility test and foreign virus test standards;

[0097] 2. Subculture and culture of production cells

[0098] After the seed batch cells SK-6 are digested and dispersed, the cells are cultured in suspension, and when they grow to a suitable density, they are used for further passage or virus inoculation;

[0099]Cell test: test according to the provisions of "Chinese Veterinary Pharmacopoeia", and the results meet the requirements of cell standards for production and inspection;

[0100] 3. Propagation of virus fluid for production

[0101] The production seed virus...

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Abstract

The invention relates to a method for producing swine fever vaccines, which comprises the following technical steps: (1) cloning and identifying hog cholera lapinized virus strain; (2) propagating seed viruses of the cloned strain; (3) propagating productive cells; (4) propagating productive virus solution; and (5) distributing vaccines, filling and freeze-drying. The method of the invention has the characteristics of simple and stable production process, high virus immunogenicity and genetic stability, small exogenous virus pollution risks, convenient examination, low production cost and thelike. The swine fever vaccines produced by the method have high genetic stability, purity and immunogenicity.

Description

technical field [0001] The invention relates to a production method of a swine fever live vaccine and belongs to the technical field of veterinary biological products. Background technique [0002] Classical Swine Fever (CSF) is a highly contagious and fatal swine infectious disease caused by Classical Swine Fever Virus (CSFV), which occurs in many countries and regions in the world. Industrial production has brought serious economic losses. The World Organization for Animal Health (OIE) lists swine fever as a statutory notifiable disease, and my country classifies it as a first-class animal infectious disease. [0003] At present, there are mainly 3 kinds of live swine fever vaccines with good effects in the world, namely the GPE strain in Japan, the Thiverval strain in France and the attenuated rabbit strain (C strain) in China, among which the attenuated rabbit strain is the most widely used and is the world's leading It has made important contributions to the control a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/04A61K39/187A61P31/14
Inventor 戴志红王在时关孚时蒋卉李翠赵耘秦玉明
Owner CHINA INST OF VETERINARY DRUG CONTROL
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