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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/bacteriophages, etc., can solve the problems of low feed conversion rate, high chicken waste rate, and difficult absorption, etc. To achieve the effect of good immunogenicity, easy absorption, and avoiding side effects

Active Publication Date: 2015-05-20
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 culture and harvest

[0016] 10-day-old SPF chicken embryos were taken to prepare primary chicken embryo fibroblasts at 3 × 10 6 Inoculation amount of cells / mL The prepared cells were inoculated into M199 medium containing 8% (v / v) fetal bovine serum, and cultured in shake flasks overnight at 37°C; when the cells were overgrown with the medium monolayer, Inoculate the cell suspension with a 0.2% (v / v) inoculation volume of the cell suspension with a titer of ≥10 6.5 TCID 50 / 0.1mL of avian reovirus, controlled temperature at 37°C and continued to culture in shake flasks for 72 hours, harvested the cell culture, and repeated freezing and thawing 3 times to obtain cytovenom containing supernatant;

[0017] 1.2 Virus inactivation

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

Embodiment 2

[0051] 1.1 Virus culture and harvest

[0052] 11-day-old SPF chicken embryos were taken to prepare primary chicken embryo fibroblasts at 3.3 × 10 6 Inoculation amount of cells / mL The prepared cells were inoculated into M199 medium containing 8% (v / v) fetal bovine serum, and cultured in shake flasks overnight at 37°C; when the cells were overgrown with the medium monolayer, Inoculate the cell suspension with the inoculation amount of 0.3% (v / v) of the cell suspension with a titer of ≥10 6.5 TCID 50 / 0.1mL of avian reovirus, controlled temperature at 37°C and continued to culture in shake flasks for 66 hours, harvested the cell culture, and repeated freezing and thawing 3 times to obtain cytovenom containing supernatant;

[0053] 1.2 Virus inactivation

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

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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 a water-in-oil-in-water type inactivated vaccine of avian reovirus. Background technique [0002] Avian reovirus is one of the main pathogens of chicken viral arthritis and tenosynovitis. It belongs to the family Reoviridae and belongs to the genus Reovirus. Since this strain was first isolated from the respiratory tract of chickens suffering from chronic respiratory disease in 1954, a variety of avian reovirus isolates have been reported successively. With the deepening of related research in recent years, it has been found that avian reovirus may cause other diseases besides viral joint and tenosynovitis. Avian reoviruses were isolated from chickens with diseases such as syndrome and temporary digestive system disorders. [0003] Avian reoviruses are distributed worldwide and are widely prevalent in chickens and turkeys. Cases of avian reovi...

Claims

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Application Information

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Patent Type & Authority Patents(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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