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Muscovy duck novel adenovirus strain, bivalent inactivated vaccine and preparation method thereof

A bivalent inactivated vaccine and adenovirus technology, applied in the field of veterinary biological products, can solve the problems of economic losses in the Muscovy duck breeding industry, and achieve good commercial development prospects and good safety effects

Active Publication Date: 2019-12-31
SHANDONG SINDER TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After the isolation and identification of the pathogen and the artificial infection test of the virus, it was preliminarily shown that the pathogen of the disease was DAdV-2 and another new type of Muscovy duck adenovirus. Some experts named this new type of adenovirus as duck adenovirus type 3 (Duck adenovirus type 3 , DAdV-3), but this nomenclature has not been accepted by the International Committee on Taxonomy of Viruses (ICTV)
Through the epidemiological investigation of the new Muscovy duck adenovirus, it was found that the disease is prevalent in almost all Muscovy duck breeding areas in my country, causing great economic losses to the Muscovy duck breeding industry
At present, there is no commercial vaccine (or antibody) for the effective prevention and treatment of Muscovy duck adenovirus type 2 and new adenopathies at home and abroad, and it is imminent to develop safe and effective vaccines and antibodies

Method used

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  • Muscovy duck novel adenovirus strain, bivalent inactivated vaccine and preparation method thereof
  • Muscovy duck novel adenovirus strain, bivalent inactivated vaccine and preparation method thereof
  • Muscovy duck novel adenovirus strain, bivalent inactivated vaccine and preparation method thereof

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Experimental program
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Embodiment 1

[0026] Virus isolation and identification

[0027] 1.1 Isolation and identification of duck adenovirus type 2

[0028] The disease materials were collected from 12-day-old Muscovy ducks in a Muscovy duck farm in Guangdong, and the liver, spleen, kidney and other tissues of the diseased Muscovy ducks were collected and grinded, and the non-toxic drug containing double antibodies was added at a ratio of 1:3 (w / v). Bacteria were homogenized in PBS, freeze-thawed 3 times, and then centrifuged at 8000r / min for 10 minutes. The supernatant was sterilized by a 0.22um filter and stored for later use. Inoculate muscovy duck embryo fibroblasts with 1% virus filtrate, culture in serum-free M199 culture medium, and culture at 37°C for 60-78 hours, obvious cytopathic changes may appear, collect virus fluid, and take supernatant to extract DNA, using the Hexon protein gene specificity, was identified by sequencing, which proved that the virus was duck adenovirus type 2, named DAdV-2-GD stra...

Embodiment 2

[0032] Preparation of DAdV-2-GD Strain Virus Seeds

[0033] Select muscovy 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 species (the third generation of the original virus species 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 passages are continuously passed, and the cytotoxicity of the 10th passage is used to inoculate the LMH cells grown to a single layer at a ratio of 1-3% of the culture medium, and the original culture medium is discarded. DMEM / F12 virus maintenance solution without bovine serum, 37°C, 5% CO 2 Under the condition of static culture for 72-96h, the virus liquid was harvested, and the freeze-thaw was repeated 1-2 times. The harvested virus liquid was continuously inoculated with LMH cells for 3 to 5 gen...

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Abstract

The invention discloses a Muscovy duck novel adenovirus strain, bivalent inactivated vaccine and a preparation method thereof. The bivalent inactivated vaccine is prepared by taking duck adenovirus and DAdV-3-GD MM virus as inactivated antigens. The Muscovy duck adenovirus type II and type III bivalent inactivated vaccine prepared from the screened virus has high safety, and any local and systemicadverse reaction caused by the vaccine does not appear. In the storage life experiment, each index is stable and effective through analysis of character, safety experiment and efficacy experiment data; and through evaluation on the immune effect of the vaccine by a serology method and a immune poison attack method, the Muscovy duck adenovirus type II and type III bivalent inactivated vaccine canprovide effective immune protection on Muscovy ducks and has good commercial development prospect.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a new muscovy duck adenovirus strain, a bivalent inactivated vaccine and a preparation method thereof Background technique [0002] Avian adenovirus is one of the common infectious disease pathogens in poultry, which can cause clinical and pathological syndromes in chickens, ducks, geese and other poultry or wild birds, including hydropericardium, hepatitis, aplastic anemia, hemorrhage, A variety of poultry conditions such as mild respiratory disease. [0003] Duck (Anas platyrhynchos domestica)-derived adenoviruses include AT-rich adenovirus genus duck-derived adenovirus A (Duck atadenovirus A, DAdV-A) and avian adenovirus genus duck-derived adenovirus B (DAdV-B). DAdV-A is duck adenovirus type 1 (Duck adenovirus type 1, DAdV-1), also known as egg drop syndrome virus (Egg drop syndrome virus, EDSV), which is related to egg drop syndrome. Egg volume drops...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/235A61P31/20C12R1/93
CPCA61K39/12A61K2039/5252A61K2039/552A61K2039/70A61P31/20C12N7/00C12N2710/10221C12N2710/10234
Inventor 罗济冠常娓娓王学波刘宁郭春丽罗展徐丽娜李朝阳
Owner SHANDONG SINDER TECH
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