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Novel chicken reovirus strain

A reovirus and vaccine technology, applied in antiviral agents, viruses/phages, viral antigen components, etc., can solve the problems of viral nucleic acid instability, base mutation, accelerated reovirus mutant strains, etc.

Inactive Publication Date: 2019-06-18
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Reovirus is a segmented double-stranded RNA virus. The viral nucleic acid is unstable and prone to base mutations, deletions, or gene recombination, resulting in the emergence of new strains
In recent years, there have been continuous reports of reovirus-caused white-feathered broiler chickens in northern my country. Non-classic reovirus strains have been isolated from chickens immunized with classical strain vaccines. These non-classi

Method used

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  • Novel chicken reovirus strain
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Embodiment 1: the isolation process of virus strain

[0014] On May 12, 2018, I received samples of the hock joints of sick chickens sent by a white-feather broiler company in Liaocheng, Shandong Province. The hock joints were opened, the joint cavity and tendons were leaked, and the exudate was dipped in with sterile cotton swabs, and scissors were used to clean the hock joints. Cut tendons and necrotic tissues, cut them into pieces and grind them, add appropriate normal saline, then add 1000U of double antibodies, freeze and thaw three times, centrifuge at 5000rpm at 4°C for 15min, absorb the supernatant, filter and sterilize with a 0.22μm filter. The above-mentioned filtrate was inoculated into 10 pieces of SPF chicken embryos through the chorioallantoic membrane, and the inoculation amount was 0.2ml / piece. After inoculation, they were placed in a biochemical incubator at 37°C for constant temperature incubation. The chicken embryos that died within 24 hours were di...

Embodiment 2

[0017] Embodiment 2: Identification of SD18 strain

[0018] 1. PCR identification

[0019] 1. Extraction and reverse transcription of viral RNA

[0020] Add 300 μl of virus chorion grinding supernatant to a 1.5ml centrifuge tube, add 700 μl Trizol Reagent RNA extraction solution, mix well, and place at room temperature for 15 minutes. Add 200 μl of chloroform, shake and mix thoroughly, and let stand at room temperature for 10 minutes. Centrifuge at 12,000 rpm for 15 min at 4°C, and transfer 500 μl of the supernatant to a 1.5 ml centrifuge tube treated with DEPC. Add 500 μl of pre-cooled isopropanol, let stand at -20°C for 30 minutes, centrifuge at 12,000 rpm for 15 minutes, and discard the supernatant. Add 1ml of 75% ethanol, centrifuge at 8000rpm for 5min, and pour off the liquid. Put it upside down on paper to dry, suspend the pellet in 30 μl of RNA-free water, and store it at -80°C for later use. The extracted RNA was reverse-transcribed according to the instruction ma...

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Abstract

The invention provides a novel chicken reovirus strain which is a chicken reovirus SD18 strain collected in CCTCC (China Center For Type Culture Collection) in Wuhan on February 20, 2019 with the collection number of CCTCC NO:V201910. The screened virus strain is used for preparing a vaccine. The chicken reovirus variant strain is obtained through pathogen separation, identification and purification of joint samples collected from clinically suspected reovirus attacked chicken flocks, the screened virus is determined to be a variant strain of reovirus popular on white feather broilers in Chinathrough RT-PCR tests, virus neutralization tests and animal regression tests, the variant strain has notable difference from classical strains in serology, and a basis is provided for comprehensive control of the disease in poultry farms in China.

Description

technical field [0001] The invention belongs to the technical field of virus screening, in particular to a novel chicken reovirus strain. Background technique [0002] Chicken reovirus (chicken reovirus, REO) is also known as "chicken viral arthritis". Chickens of various breeds and ages can be infected with the disease, among which the northern white-feathered broiler chickens are the most common. It can spread vertically, resulting in a great reduction in production performance and seriously endangering the development of the chicken industry. Avian reoviruses are ubiquitous and affect a wide variety of poultry, including chickens, ducks, and geese, causing enormous economic losses, especially in broiler chickens causing viral arthritis, respiratory disease, hepatitis, myocarditis, immunosuppression, and stunting syndrome and other clinical symptoms. [0003] Reovirus is a segmented double-stranded RNA virus. The nucleic acid of the virus is unstable, and it is prone to ...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/15A61P31/14
Inventor 杜元钊张玉杰刘东刘红祥范根成
Owner YEBIO BIOENG OF QINGDAO
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