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Goose astrovirus variation strain

An astrovirus and inactivated vaccine technology, applied in the field of vaccine virus screening, can solve problems such as economic loss, gosling disease, and threats to the development of the goose industry.

Active Publication Date: 2020-05-15
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since around 2010, a highly fatal visceral gout has occurred in the vast goose-raising areas of our country. The disease spreads rapidly and often causes all the goslings hatched in the same batch to get sick. The epidemic has caused huge economic losses and seriously threatened the development of my country's goose industry.

Method used

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  • Goose astrovirus variation strain
  • Goose astrovirus variation strain
  • Goose astrovirus variation strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: the isolation process of virus strain

[0023] From October 2017 to May 2019, 5-15 day-old sick and dead goslings sent for inspection in Guangdong, Sichuan, Hebei, Shandong, Anhui, Liaoning and other places, the goslings appeared with swollen hocks, toes and knuckles, white free Urate deposits, sick geese like to crouch, bi-webs dare not touch the ground, eyes closed and lethargic, and the whole body trembles involuntarily. Autopsy showed severe gout in the whole body organs, and white urate deposits in the heart, liver, and kidneys. In some cases, urate deposits were also found on the inner surface of the glandular stomach, subcutaneously, and in the gallbladder.

[0024] Aseptically take an appropriate amount of liver and kidney of dead goslings, add 5 times the volume of sterile normal saline, cut into pieces, fully homogenate, centrifuge at 8000g for 15min, take the supernatant and filter it through a 0.22μm filter to sterilize, pass through the chorio...

Embodiment 2

[0027] Embodiment 2: Identification of goose astrovirus FLX variant virus SC-3 strain

[0028] 1. Transmission electron microscope observation

[0029] Take 200ml of allantoic fluid from F3 goose embryos that has been blindly passed, centrifuge at 10,000×g for 10 minutes to remove larger impurities, then centrifuge at 40,000×g for 3 hours at 4°C to further remove smaller impurities, and resuspend the virus particles in 60μl PBS , negatively stained with 1% phosphotungstic acid for 30s, and observed the virus particle structure under a transmission electron microscope.

[0030] The diameter of the virus particles in the electron microscope field of view is about 30nm, and the outer surface of the virus has no envelope and is spherical, which conforms to the structural characteristics of astroviruses ( figure 2 ).

[0031] 2. PCR identification

[0032] 1. Extraction and reverse transcription of viral nucleic acid

[0033] Add 300 μl of virus chorion grinding supernatant to...

Embodiment 3

[0048] Embodiment 3, preparation of vaccine and virus neutralization test

[0049] 3.1 Preparation of vaccine

[0050] 3.1.1 Propagation of goose astrovirus The classic goose astrovirus YB-1 strain (YB-1 strain belongs to the classic goose astrovirus FLX branch, and its LMH cell-adapted strain was isolated by Qingdao Yibang Bioengineering Co., Ltd. identification and preservation) inoculated on the LMH cells covered with a single layer, observed continuously for 5 to 6 days, and reproduced the required antigens; the goose astrovirus FLX variant virus SC-3 strain virus screened by the present invention was made with sterilized physiological saline 100-fold dilution, inoculate 10-day-old goose embryos through the chorioallantoic membrane, 0.2ml per embryo, seal the pinhole after inoculation, and continue incubation at 37°C without turning the embryos. The embryos were photographed once a day, and the goose embryos that died within 48 hours were discarded. Thereafter, the embry...

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Abstract

The invention provides a goose astrovirus variation strain. The virus is a variation strain of classical goose astrovirus FLX, and the preservation number is CCTCC NO: V201976. The screened virus strain is applied to preparation of vaccine. Isolation, identification and purification of pathogens is performed on clinical gosling gout samples so as to obtain the goose astrovirus strain, and it is confirmed through observation of a transmission electron microscope, RT-PCR tests, serological tests and animal regression tests that the screened virus is the variation strain of the classic goose astrovirus FLX; and the variation strain has significant differences of serological and biological features from the classic strain, and a basis is provided for comprehensive prevention and control of thedisease in goose farms in China.

Description

technical field [0001] The invention belongs to the technical field of vaccine virus screening, and in particular relates to a mutant strain of goose astrovirus. Background technique [0002] Goose Astrovirus is a single-stranded positive-strand, non-enveloped RNA virus with a full-length viral genome of 7.9kb. The virus is one of the main pathogens causing diarrhea in animals. In addition to causing diarrhea and poor growth, it also infects other tissues and organs of the body, resulting in fatal hepatitis, nephritis, and goose visceral gout. Since around 2010, a highly fatal visceral gout has occurred in the vast goose-raising areas of our country. The disease spreads rapidly and often causes all the goslings hatched in the same batch to get sick. The epidemic has caused huge economic losses and seriously threatened the development of goose industry in China. The disease is mainly characterized by a large amount of urate deposits covering the viscera, joints and muscles....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N7/06A61K39/12A61P31/14
CPCC12N7/00A61K39/12A61P31/14C12N2770/12021C12N2770/12034A61K2039/552
Inventor 张玉杰刘东刘红祥
Owner YEBIO BIOENG OF QINGDAO
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