Process for preparing subunit vaccine of reovirus antigen for grass carp

A reovirus and subunit vaccine technology, applied in the field of virology, can solve the problems of limited application and promotion value, and achieve the effects of high packaging volume, convenient operation, and exclusion of genetic contamination

Inactive Publication Date: 2005-05-18
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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  • Abstract
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AI Technical Summary

Problems solved by technology

However, due to many unfavorable factors in the stability, purity and safety of inactivated vaccines, their application and promotion value is limited.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0012] The grass carp reovirus antigen subunit vaccine provided by the invention comprises:

[0013] (1) CIK cells (grass carp kidney cell line) were used for virus propagation, and the virus suspension of grass carp reovirus Hunan Shaoyang strain GCRV873 infected with CIK cells was obtained.

[0014] (2) Purify and concentrate the cell virus suspension infected with GCRV873 by centrifugation.

[0015] (3) The purified virus is treated with a surfactant to disintegrate the complete virus particle structure to form viral nucleic acid and capsid protein subunit components. Surfactant can adopt NP40 (Fluka company product); Its concentration is 0.5%~1%, can make virus capsid protein become loose like this and will not degrade completely.

[0016] (4) Nuclease is used for digestion to degrade the free viral genome dsRNA in the GCRV873 subunit preparation.

[0017] (5) A nucleic acid-free grass carp reovirus antigen subunit protein preparation is obtained by dialysis treatment. ...

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PUM

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Abstract

The preparation process of reovirus antigen subunit vaccine for grass carp includes the following steps: proliferation of GCRV873, which is the Hunan Shaoyang strain of reovirus, in grass carp kidney cell line; centrifugal purification and concentration of GCRV873 infected cell virus suspension; processing purified virus with surfactant to disintegrate complete virus structure to form viral nucleic acid and capsid protein subunit component; digesting with nuclease to degrade free virus genome dsRNA; dialysis to obtain reovirus protein subunit antigen preparation for grass carp containing no nuclease; and diluting with physiological saline to obtain the said vaccine. The vaccine of the present invention has the advantages of containing complete virus protein antigen component, containing no genetic virus matter RNA, simple preparation process, high product purity, etc.

Description

technical field [0001] The invention relates to the field of virology, in particular to a method for preparing a grass carp reovirus antigen subunit vaccine. The virus strain used in the present invention is grass carp reovirus Hunan Shaoyang strain GCRV873 (Grass carp reovirus), also known as grass carp hemorrhagic disease virus Hunan Shaoyang strain GCHV873 (Grass carp hemorrhagic virus), has been preserved by the China Center for Type Culture Collection, and the preservation date It was dated July 23, 2004, and the deposit number is CCTCC NO: V200414. Background technique [0002] Grass carp (Ctenopharyngodon idellus) is one of the four major freshwater fish species in my country. Grass carp reovirus is the main pathogen that causes the death of grass carp. When the disease is severe, it can cause more than 85% of grass carp fry to die, thus causing serious economic losses to the freshwater aquaculture industry. For the prevention and treatment of viral diseases, vaccin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61P31/12
Inventor 方勤丁清泉
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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