Preparation method of inactivated vaccine for iridovirus of grouper

A technology of iridescent virus and inactivated vaccine, which is applied in antiviral agents, virus antigen components, etc., can solve the problem of not seeing grouper iridescent virus inactivated vaccine, etc., and achieve good immune protection effect, simple equipment requirements, and inactivated vaccines. The effect of short life time

Active Publication Date: 2011-09-14
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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AI Technical Summary

Problems solved by technology

Inactivated vaccines have strong safety and immunogenicity. In mammals and birds, there have been successful precedents of large-scale propagation of viruses and production of inactivated virus vaccines using cell lines. There is no report on the use of cell lines to prepare grouper iridescent virus inactivated vaccines

Method used

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  • Preparation method of inactivated vaccine for iridovirus of grouper
  • Preparation method of inactivated vaccine for iridovirus of grouper

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Embodiment 1

[0014] 1. Cultivation of grouper embryonic cells for amplification of grouper iridescent virus. Take grouper embryo cells covered with a single layer, remove the original culture medium, rinse with phosphate buffered saline solution, add 0.25% trypsin to digest for 2 minutes, add the newly prepared MEM culture medium containing 10% fetal bovine serum (pH7 .4), after blowing evenly, divide it into 3 cell culture flasks for culture and expansion at 25°C, and inoculate grouper iridescent virus amplification when the cells grow to the logarithmic phase.

[0015] 2. Amplification and harvest of grouper iridescent virus. Inoculate 0.05 MOI (multiplicity of infection, multiplicity of infection) virus in logarithmic phase cells and culture in a 25°C incubator to amplify the virus. After 4 days of continuous culture, the cytopathy was complete. After repeated freezing and thawing 3 times, divide into 50 ml centrifuge tubes, centrifuge at 2500g for 10 minutes to remove cell debris, col...

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Abstract

The invention relates to a preparation method of an inactivated vaccine for the iridovirus of a grouper, comprising the following steps of: inoculating the iridovirus to embryonic cells of the grouper in a logarithmic phase by taking an embryonic fine system of the grouper as an amplification system of the iridovirus; repeatedly freezing and thawing and centrifugalizing after complete lesion, and inactivating an obtained iridovirus solution at 4 DEG C for 16 hours by using beta-propiolactone with the final concentration of 1:500 so as to obtain the inactivated iridovirus vaccine. The inactivated iridovirus vaccine is applied to a juvenile immunized Malabar grouper, and an iridovirus counteracting result shows that relative protection ratio is more than 90 percent after 15 days. The preparation method has the advantages of easiness and convenience for operation, simple equipment requirement and good repeatability and keeps good immunogenicity of the iridovirus under the precondition of efficiently inactivating the iridovirus, thereby having good immune protection effect; in addition, the invention can be used for the preventive immune of the grouper, thereby enhancing the survival rate and the culturing efficiency of cultured groupers.

Description

technical field [0001] The invention belongs to the technical field of new veterinary biomedicine, and in particular relates to a method for producing grouper iridescent virus inactivated vaccine by using fish cells. Background technique [0002] Grouper meat is delicious and has high protein content. It is a kind of edible fish with high economic value, and it is also an excellent fish species that earns foreign exchange in my country. However, in recent years, with the expansion of breeding scale and the deteriorating breeding environment, various diseases, especially viral diseases, frequently broke out, causing significant economic losses. Iridescent virus is an important pathogen of grouper, which can cause fish mortality ranging from 30% (in adult fish) to 100% (juvenile stage). In Asia, especially when the water temperature is above 22°C, this virus is very easy to infect rare seawater fish cultured in China, Japan and Southeast Asia, such as grouper, barramundi, red...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61P31/20
Inventor 秦启伟欧阳征亮汪沛然黄晓红黄友华
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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