Preparation method for avian reovirus virus water-in-oil-in-water type inactivated vaccines

A technology of avian reovirus and water-in-oil-in-water, which is applied in the fields of biochemical equipment and methods, antiviral agents, viruses/phages, etc., can solve the problems of low feed conversion rate, high chicken waste rate, economic loss, etc. To achieve the effect of good immunogenicity, easy absorption, and avoiding side effects

Active Publication Date: 2014-01-01
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Arthritis, tenosynovitis and other diseases caused by avian reovirus will cause high abandonment rate, growth inhibition and low feed conversion rate of chickens. Therefore, the prevalence of avian reovirus often causes huge economic losses, especially for In the environment where poultry are concentrated in chicken farms, if a large-scale outbreak of avian reovirus occurs, it will become difficult to control
[0004] In the prior art, the main means to control the incidence of avian reovirus is to inoculate inactivated whole virus vaccine, and to control typical chicken reovirus infection through immunization, but most of the inactivated vaccines are water-in-oil type at present, and injection The time resistance is large, it is not easy to absorb, and has certain toxic and side effects

Method used

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  • Preparation method for avian reovirus virus water-in-oil-in-water type inactivated vaccines
  • Preparation method for avian reovirus virus water-in-oil-in-water type inactivated vaccines
  • Preparation method for avian reovirus virus water-in-oil-in-water type inactivated vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] 1.1 Virus cultivation and harvest

[0016] Take 10-day-old SPF chicken embryos to prepare primary chicken embryo fibroblasts, and use 3×10 6 Inoculation amount of cells / mL The prepared cells were inoculated in M199 medium containing 8% (v / v) fetal bovine serum, and cultured overnight in shake flasks at 37°C; when the cells covered the monolayer of the medium, According to the inoculum amount of 0.2% (v / v) of the cell suspension, inoculate the cell suspension with a toxicity value ≥ 10 6.5 TCID 50 / 0.1mL of avian reovirus, control the temperature at 37°C and continue to shake the flask for 72 hours, harvest the cell culture, freeze and thaw repeatedly 3 times, and obtain the cell venom containing the supernatant;

[0017] 1.2 Inactivation of virus

[0018] Put the cell venom containing the supernatant obtained above in a sterilized container, add formaldehyde solution according to the volume of the virus liquid and adjust the final concentration of formaldehyde to 1‰ ...

Embodiment 2

[0051] 1.1 Virus cultivation and harvest

[0052] Take 11-day-old SPF chicken embryos, prepare primary chicken embryo fibroblasts, and use 3.3×10 6 Inoculation amount of cells / mL The prepared cells were inoculated in M199 medium containing 8% (v / v) fetal bovine serum, and cultured overnight in shake flasks at 37°C; when the cells covered the monolayer of the medium, According to the inoculum amount of 0.3% (v / v) of the cell suspension, inoculate the cell suspension with a toxicity value ≥ 10 6.5 TCID 50 / 0.1mL of avian reovirus, control the temperature at 37°C and continue to shake the flask for 66 hours, harvest the cell culture, freeze and thaw repeatedly 3 times, and obtain the cell venom containing the supernatant;

[0053] 1.2 Inactivation of virus

[0054] Put the cell venom containing the supernatant obtained above in a sterilized container, add formaldehyde solution according to the volume of the virus liquid and adjust the final concentration of formaldehyde to 0.8...

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Abstract

The invention provides a preparation method for avian reovirus virus water-in-oil-in-water type inactivated vaccines. The preparation method comprises the following steps: preparation of chicken embryo fibroblast, cultivation of viruses, inactivation of the viruses, emulsification of the vaccines, and the like. Compared with the prior art, locally separated avian reoviruses are adopted, so that the immunogenicity is good, and the relatively high protection rate is adopted; the chicken embryo fibroblast is adopted to replace the utilization of a chicken embryo chorioallantoic membrane in a inoculation method for breeding the avian reoviruses, so that the vaccine preparation cost is saved, the mass discrepancy between each batch is lowered, the yield is high, and the viral titer and the vaccine quality are improved; the vaccines are subjected to emulsification by using a homogenizer, the prepared vaccines are water-in-oil-in-water type; compared with traditional water-in-oil type vaccines, the vaccines provided by the invention have low viscosity, small granularity, and easy absorption; surface active agent compositions are not required to be added in the preparation method by selecting and using an adjuvant, so that the prepared vaccines have good safety, and avoid side effects caused by difficult absorption of a traditional oil adjuvant.

Description

technical field [0001] The invention relates to the technical field of veterinary vaccines, in particular to a method for preparing an avian reovirus-inactivated water-in-oil-in-water vaccine. Background technique [0002] Avian reovirus is one of the main pathogens of chicken viral arthritis and tenosynovitis, belonging to Reoviridae, a kind of Reovirus genus. Since the strain was first isolated from the respiratory tract of chickens suffering from chronic respiratory diseases in 1954, a variety of avian reovirus isolates have been reported one after another. With the deepening of relevant research in recent years, it has been found that avian reovirus may cause other diseases besides viral joints and tenosynovitis. Avian reovirus was isolated from chickens with diseases such as syndrome and temporary digestive system disorders. [0003] Avian reovirus is distributed all over the world and is widely prevalent in chickens and turkeys. Cases of avian reovirus infection in o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/15A61K9/113A61P31/14C12N7/00C12R1/93
Inventor 张立霞康亚男王寿山毛雅元何召庆杨保收梁武李守军
Owner TIANJIN RINGPU BIO TECH
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