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Swine fever live vaccine ST cell culture method

A swine fever live vaccine and cell culture technology are used in the preparation of swine fever live vaccines and the field of preparation of swine fever attenuated vaccines to achieve the effects of facilitating cell dispersion, good safety and avoiding bacterial contamination

Inactive Publication Date: 2009-07-29
RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 2. Chronic swine fever
[0006] 3. Atypical swine fever
However, because the IB-RS-2 cell line can spontaneously attenuate during the process of continuous inoculation and passage with HCV, and in recent years, through nested PCR detection, the parent of this cell line has already been contaminated by an antigen-variant HCV, so ATCC has This cell line was destroyed in 1994

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1: Inoculation and subculture of swine fever HCV virus

[0037]1. The incubation period for collection is 48-96 hours, the body temperature rises in an obvious curve, at least 3 times the temperature exceeds the normal temperature by more than 1°C, and the rabbit spleen with stereotyped fever is retained for 18-36 hours as the virus species.

[0038] 2. Take the ST cell spinner bottle that has formed a good monolayer, discard the nutrient solution, grind the spleen toxin at a concentration of 0.2% to 0.3% of the cell spleen toxin inoculum, inoculate it on the ST cells, add the cell growth solution, and place Continue culturing at 36-37°C.

[0039] 3. Harvest and change the medium for the first time 5 days after inoculation, and then harvest and change the medium every 4 days.

[0040] 4. The harvested virus liquid is subjected to sterility test, virus content determination, and after the safety test is qualified, 5% sucrose skim milk is added to stabilize ec...

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Abstract

The invention provides a culture method for a novel swine fever living vaccine cell roller bottle, which comprises the steps as follows: the spleen of a rabbit having stereotyped thermal reaction is collected as a virus seed by rabbit body immunity HCV; an ST monolayer cell is prepared, added with cell culture liquid and arranged into a roller bottle incubator of 37 DEG C for culture; the collected rabbit spleen having stereotyped thermal reaction is inoculated on the ST monolayer cell at a concentration of 0.2 to 0.3 percent; then the cell culture liquid is added and cultured in the roller bottle incubator of 37 DEG C; virus liquid 1 is collected after the mixture is cultured for 5 days; and then the virus liquid is collected every 4 days; the collected virus liquid is frozen and dried in vacuum, thus becoming the swine fever living vaccine. The invention aims at overcoming the defect of lower culture titre of the original cell line to HCV and can be used for improving the antigenicity of the swine fever living vaccine and enhancing the immunity of an organism.

Description

technical field [0001] The invention relates to a preparation method of a live swine fever vaccine, specifically, the attenuated swine fever vaccine is prepared by cultivating the swine fever virus in a transfer bottle of subculture cells, that is, ST cells. Belongs to the field of bioengineering. Background technique [0002] Classical swine fever (HC) is a highly contagious disease of pigs. It spreads all over the world and causes huge damage to the economy. Classical swine fever virus (CSFV) is a highly contagious infectious disease caused by classical swine fever virus (CSFV). It is characterized by strong infectivity and high lethality. Pigs of different ages, sexes and breeds can be infected. One of the major infectious diseases closely watched by the World Food and Agriculture Organization and governments of various countries. In the International Committee's "International Sanitary Code" (2002 edition), swine fever is listed as one of the 15 legal infectious diseas...

Claims

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Application Information

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IPC IPC(8): C12N7/00
Inventor 张桂红杨保收
Owner RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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