Duck adenovirus type 2 inactivated vaccine

A technology of inactivated vaccines and adenoviruses, applied in vaccines, viruses, antiviral agents, etc., can solve the problem of lack of prevention, and achieve the effects of good safety, good commercial development prospects, and effective immune protection.

Active Publication Date: 2017-09-08
SHANDONG SINDER TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no commercial inactivated vaccine to prevent the disease at home and abroad, and the disease is still spreading

Method used

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  • Duck adenovirus type 2 inactivated vaccine
  • Duck adenovirus type 2 inactivated vaccine
  • Duck adenovirus type 2 inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Isolation and identification of GD strain

[0016] 1. Epidemiological investigation Since 2014, a group I duck type 2 adenovirus disease with high mortality has appeared in some muscovy ducks in Guangdong, Zhejiang and other regions. After the autopsy of the muscovy duck that died of illness, the main pathological changes were as follows: hepatomegaly, hepatic necrosis, splenomegaly, hemorrhagic spots, pancreas necrosis and other diseases characterized by multiple organ lesions. After clinical investigation and laboratory testing, the preliminary diagnosis was Group I duck adenovirus type 2. In 2015, the inventor successfully isolated a virus from ducks in a duck farm in Guangdong.

[0017] 2. Virus isolation Take 50-100 g of liver, spleen, pericardial fluid, pancreas, and bursa of the diseased duck, grind the tissue with a mortar, and add liquid nitrogen continuously until it is ground into powder (no obvious visible particles) Homogenize with sterile normal saline a...

Embodiment 2

[0033] Preparation of GD Strain Virus Seeds

[0034] Select the duck embryo fibroblasts that have grown into a single layer, discard the original culture medium, add serum-free M199 maintenance solution with a final concentration of 1% virus seed (the third generation of the original virus strain of GD strain), and culture at 37°C for 60~ After 78 hours, obvious cytopathy can appear, and harvest when the cytopathy reaches more than 80%. According to this method, 10 generations were passed continuously, which were respectively marked as C3-C10 generations. The virus content of each passage was determined. Will test for sterility and virus content ≥10 6.0 TCID 50 The virus liquid is mixed, quantitatively divided, and stored in a freeze-dried manner.

Embodiment 3

[0036] Preparation of GD strain antigen

[0037] 1 Selection of seedling materials Preparation of duck embryo fibroblasts and GD strain virus liquid.

[0038] 2. Cell preparation Take 11-13 day-old SPF duck embryos, digest them with 0.25% trypsin, add an appropriate amount of M199 culture medium containing 10% serum for culture.

[0039] 3 Inoculation of virus Select the duck embryo fibroblasts that have grown into a single layer, discard the original culture medium, add the maintenance solution with a final concentration of 1% virus species, and place in a 37°C incubator to continue culturing.

[0040] 4 Harvesting After inoculation, observe twice a day and record the cell lesions. Harvest when the cytopathic rate reaches more than 80%, freeze and thaw twice, and store the harvested cytotoxicity at -20°C before inactivation.

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Abstract

The invention aims at providing a duck adenovirus type 2 inactivated vaccine, wherein an antigen is an inactivated GD strain virus; and the GD strain virus is preserved in China Center for Type Culture Collection in Wuhan University on June 5, 2016 with preservation number of CCTCC No: V201633. The duck adenovirus type 2 inactivated vaccine prepared by the invention is good in safety and free from any local and systemic adverse reactions caused by the vaccine. Based upon analysis on characters, safety test and efficacy test data in a preservation period test, various indexes are stable and effective; and a result of assessing an immune effect of the vaccine by virtue of a serological method and an immune attack method shows that the inactivated vaccine prepared by the invention can achieve effective immune protection on ducks, and the inactivated vaccine has a good commercial development prospect.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, in particular to an inactivated duck type 2 adenovirus vaccine. [0002] technical background [0003] Adenoviruses are common infectious pathogens of poultry. Avian adenoviruses are divided into three groups: Group I is avian adenoviruses isolated from respiratory infections in chickens, turkeys, geese and ducks. They have common group-specific antigens and can be divided into A, B, C, D, E5 species and 12 serotypes; Group II includes turkey hemorrhagic enteritis virus, pheasant marble skin disease virus and chicken large spleen disease virus; Group III is an adenovirus isolated from chicken laying syndrome and ducks. Duck adenovirus type 2 belongs to Group I poultry adenovirus. It was isolated from sick Muscovy ducks by French-Hungarians in 1977. The infected ducks mainly manifested symptoms such as depression, diarrhea, and emaciation. Once the virus is brought into the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/235A61P31/20C12N7/00
CPCA61K39/12A61K2039/5252A61K2039/552C12N7/00C12N2710/10221C12N2710/10234
Inventor 王相芹王丹娜刘阳李晓林张海波胡秀香李焕明林文超张玉杰谭鹏邵振文李明义李朝阳
Owner SHANDONG SINDER TECH
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