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Method for Propagating Infectious Bursal Virus with Chicken Embryo Origin Cell Line to Prepare Inactivated Vaccine and Combined Vaccine

A technology for bursal disease and inactivated vaccine, which is applied in the direction of antiviral agents, viral antigen components, and medical preparations containing active ingredients, etc., which can solve the problem of low titer of cultured virus, increase production cost, and increase concentration multiple. and other issues to achieve the effect of improving immune efficacy, saving production costs and ensuring safety.

Inactive Publication Date: 2011-11-30
POULTRY DISEASE RES INST OF HENAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The spinner bottle primary cell (fibroblast) culture method is that the cultured cells adhere to the wall and grow on the inner wall of the glass culture flask. The technology is mature and stable, but the cultured virus titer is not high, usually at 10 -6.0～6.5 , and the batch-to-batch difference is large, it is necessary to increase the concentration multiple and increase the production cost when making seedlings
In addition, a large number of chicken embryos are used in production, which is not only labor-intensive, but also has potential safety hazards of exogenous virus contamination.

Method used

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  • Method for Propagating Infectious Bursal Virus with Chicken Embryo Origin Cell Line to Prepare Inactivated Vaccine and Combined Vaccine
  • Method for Propagating Infectious Bursal Virus with Chicken Embryo Origin Cell Line to Prepare Inactivated Vaccine and Combined Vaccine
  • Method for Propagating Infectious Bursal Virus with Chicken Embryo Origin Cell Line to Prepare Inactivated Vaccine and Combined Vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Isolation, cultivation and identification process of infectious bursal virus HQ strain

[0030]In the early 1990s, the super-virulent infectious bursa virus that was prevalent in the world was introduced into China, causing a large number of morbidity and death in unimmunized chicks, with a mortality rate as high as 40-60%. The inventor collected the bursa of dead chickens in June, 1992 in a diseased chicken farm in the suburbs of Zhengzhou for virus isolation. HQ strain cystic virus is a super-virulent strain isolated from chickens with outbreaks of infectious bursal disease. After subculture and training, the pathogenicity of cytotoxicity is weakened after training and training, and the adaptability to cells is greatly enhanced. The toxic price on fibroblasts, TCID 50 ≤10 -6.0 / 0.1ml; toxicity on DF-1 cells, TCID 50 ≤10 -7.0 / 0.1ml; HQ strain cytotoxicity was cloned and purified and tested for exogenous diseases. In early 2000, the China Veterinary Drug ...

Embodiment 2

[0031] Example 2 Isolation, cultivation and identification process of egg drop syndrome virus Z16 strain

[0032] In the early 1990s, Egg Drop Syndrome (EDS) broke out abroad as early as 1976 -76 ) disease was introduced into China, causing a large number of laying hens to fall ill. The disease has no obvious clinical symptoms, but egg production has decreased significantly, and white shells, soft shells, and deformed eggs have greatly increased. The inventor took samples from the fallopian tubes of diseased chickens in a chicken farm in the suburbs of Zhengzhou in May 1993, and continuously blindly passed 4 to 5 generations on 12-day-old duck embryos, and the duck embryos died regularly, and blood appeared in the embryo fluid. Coagulation potency. As the number of passages increases, the hemagglutination value gradually rises to the maximum value, and the HA titer can reach more than 20,000 to 80,000. After being cloned and purified by the limited dilution method, it was s...

Embodiment 3

[0033] Embodiment 3 A kind of preparation method of infectious bursal disease inactivated single vaccine comprises the following steps:

[0034] (1) Passage and culture of cells for seedling production: select chicken embryo-derived passage cell line DF-1, digest and passage with EDTA-trypsin cell dispersion liquid, continue to culture with cell growth liquid, and when a good monolayer is formed, use for Continue to subculture or inoculate the virus; the formula of the cell growth medium is: DMEM / F12 (GIBCO 12500) containing 10% superior fetal bovine serum (Wuhan Sanli), add appropriate amount of antibiotics (100U / ml), adjust the pH to 7.0-7.2 ;

[0035] (2) Propagation of cytotoxic species: take the toxic species for production, inoculate the chick embryo subculture cell line that has grown into a good single layer according to 1% of the maintenance liquid volume, and continue to cultivate at 37°C, and the cytopathic rate reaches more than 75%. Harvesting the cell fluid at t...

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Abstract

The invention relates to a method for preparing a vaccine by breeding infectious bursal disease virus (IBDV) by a chicken embryo source cell line. The method mainly comprises the following steps of: 1) subculturing cells DF-1 for preparing vaccines; 2) breeding IBDV HQ cell seeds; 3) breeding virus liquid for preparing the vaccines; 4) concentrating and inactivating the virus liquid for preparingthe vaccine; 5) preparing other virus liquid of newcastle disease method, newcastle disease-infectious bronchitis virus method, and newcastle disease-infectious bronchitis virus-egg drop syndrome method combined vaccine and concentrating; and 6) proportioning inactivated combined vaccine, emulsifying and sub-packaging. The production process is simple, and stable and is easy to operate, eliminates biological potential safety hazard existing in the conventional vaccine production, and overcomes the defects that large-scale production of the vaccines is limited by supply of chicken embryos; cost and batch-to-batch variation are reduced; the virus titer and the quality of vaccine are improved; basis is laid for culturing virus liquid on large scale by a suspension culture technology in vaccine industry; and the produced IBDV inactivated vaccine and combined vaccine have high safety and immune efficacy, and have the complete immune protection effect on IBDV attack.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a method for propagating infectious bursal virus with chicken embryo-derived cell lines to prepare inactivated vaccines and combined vaccines. Background technique [0002] Infectious Bursal Disease Virus (IBDV) belongs to the double-stranded RNA virus family and belongs to the double-stranded RNA virus genus. The pathogen mainly destroys the central immune organ of chickens - the bursa of Fabricius, causing the body's immunosuppression, leading to the death of infected chicks and the increase of infectivity to (other) pathogenic factors, and the decline of immune response to (other) vaccines. cause serious harm. At present, the disease is mainly prevented and controlled through vaccination. In addition to immunizing chicks with live vaccines, breeders are usually injected with inactivated vaccines to obtain high maternal antibodies to protect them from i...

Claims

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Application Information

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IPC IPC(8): A61K39/12A61K39/295A61P31/14A61P31/20A61K39/17A61K39/215A61K39/235
Inventor 王泽霖王川庆赵军陈陆王新卫杨霞常洪涛姚惠霞刘红英王雷王永生
Owner POULTRY DISEASE RES INST OF HENAN AGRI UNIV
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