Duck adenovirus type 2 strain

An adenovirus and disease technology, applied in the field of duck adenovirus type 2 strains, 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

Method used

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  • Duck adenovirus type 2 strain
  • Duck adenovirus type 2 strain
  • Duck adenovirus type 2 strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Isolation and identification of GD strain

[0014] 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.

[0015] 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

[0031] Preparation of GD Strain Virus Seeds

[0032] 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 solution is mixed, quantitatively divided, and stored in a freeze-dried manner.

Embodiment 3

[0034] Preparation of GD strain antigen

[0035] 1 Material selection for seedling preparation Duck embryo fibroblasts and preparation of GD strain virus liquid.

[0036] 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.

[0037] 3 Inoculation of virus Select the duck embryo fibroblasts that have grown to 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.

[0038] 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, wherein the duck adenovirus type 2 is a duck adenovirus type 2 GD strain which 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 prepared by the invention can be used for preparing a vaccine for preventing adenovirus type 2 diseases in a group of ducks. The duck adenovirus type 2 inactivated vaccine prepared by the virus screened 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 the 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 a duck type 2 adenovirus strain. [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 duck flock, i...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/235A61P31/20C12R1/93
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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