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Avian infectious bronchitis virus natural mutant

A technology of natural mutation and bronchitis, applied in the field of virology, can solve the problems of not being able to protect chickens well against the attack of wild virus and the difficulty of prevention and control

Inactive Publication Date: 2016-06-01
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the emergence of IBV mutant strains, the existing vaccines cannot protect chickens well against the attack of wild viruses, and outbreaks of IB in chicken farms occur from time to time, which brings great difficulties to the prevention and control of IBV

Method used

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  • Avian infectious bronchitis virus natural mutant
  • Avian infectious bronchitis virus natural mutant
  • Avian infectious bronchitis virus natural mutant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] [Example 1] JH06111 isolation and identification

[0029] 1 Virus isolation and identification

[0030] In 2006, the virus was isolated from the suspected IBV-infected chicken kidney tissue collected in Jinhua, Zhejiang. Specific method: dissect the sick and dead chickens for inspection, and collect kidney tissue. After the kidney tissue was cut into small pieces, sterile PBS was added at a ratio of 1:3 to 1:10, thoroughly ground, and repeatedly frozen and thawed at -70°C and 37°C three times. After centrifugation at 5000rpm for 5 minutes, the supernatant was taken and sterilized by filtration with a 0.22 μm filter membrane. Inoculate 9-11-day-old SPF chicken embryos through the allantoic cavity of chicken embryos at a dose of 0.2 mL / embryo, culture at 37°C for 72 hours, collect allantoic fluid, and pass blindly for 3 generations. Blind transfer to the third generation of chick embryos was observed to curl up, similar to typical dwarf embryos. The allantoic fluid of...

Embodiment 2

[0033] [Example 2] JH06111 pathogenicity and the detection of infected chicken virus

[0034] The allantoic fluid of chicken embryos infected with the JH06111 whose gene copy number has been determined above was treated with IBVM gene 4*10 6 Copies / 200ul / capacity was used to inoculate 10 1-day-old SPF chicks with nasal drops, and sterilized PBS-inoculated chickens were used as negative controls. The inoculated chicks were reared for 30 days in a negative-pressure high-efficiency polymer air-filtered stainless steel isolator. Observe the clinical symptoms of the chickens every day, including mental state, food intake, whether breathing is smooth, and whether the feces are normal. 5-7 days after inoculation, 2 chickens were autopsied to observe the lesions. On the 3rd day after JH06111 inoculation, the test chickens had mild depression and dyspnea, and no death occurred during the test. The autopsy found that the kidneys had no obvious swelling and no urate deposition.

[00...

Embodiment 3

[0036] [Example 3] JH06111 infected chicken antibody level detection and challenge test

[0037] Take IBVM gene 4*10 6 Copies / 200ul / capacity was used to inoculate 10 14-day-old SPF chicks with nasal drops, and sterilized PBS-inoculated chickens were used as negative controls. The blood was collected the next day after inoculation, and the antibody level in the serum of vaccinated chickens was detected by the IBVnsp5-ELISA method established in the laboratory. As a result, on the 9th day after inoculation, it was detected that the seroantibody conversion of the inoculated chickens continued until 27 days after the inoculation, and the antibody also maintained a higher level ( image 3 ). In order to understand whether the existing serum antibodies are protective, 30 days after inoculation, the IBV virulent strain JH06011 was used to challenge the virus. As a result, all chickens vaccinated with JH06111 survived without obvious clinical symptoms and autopsy changes, while the ...

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Abstract

The invention relates to the field of virology and aims to provide an avian infectious bronchitis virus natural mutant, kept in China General Microbiological Culture Collection Center, named infectious bronchitis virus and collected under CGMCC No. 11491. A genome of this mutant is 27, 541 lbp in length, a complete genome sequence of the mutant is as shown in SEQ ID NO: 1, with structural characteristics of a typical IBV genome coding structure. This mutant having special genes inserted in and provided by the invention can break the immunity protection of vaccines to breed in an organism, this mutant can proliferate in susceptible chickens without causing significant pathogenicity to sensitive chickens, and thus this mutant can serve as base material for the further study on mutation mechanism of an attenuated vaccine or IBV virus strain.

Description

technical field [0001] The invention belongs to the field of virology and relates to a natural mutant strain of infectious bronchitis virus. Background technique [0002] Chicken infectious bronchitis (Infectiousbronchitis, IB) is an acute, highly contagious infectious disease of chickens caused by infectious bronchitis virus (Infectiousbronchitis Virus, IBV). Characterized by dyspnea, nephritis and decreased egg quality, the disease is widespread worldwide. Chicken infectious bronchitis is highly contagious in chicken flocks. It is an important respiratory disease in chicken farms without highly pathogenic avian influenza and rapid Newcastle disease, and it is very harmful to poultry farming. [0003] IBV belongs to the Nidovirilaes family Coronaviridae subgenus Coronavirus gamma coronavirus subgenus, is the representative virus of this subgroup, and is also the earliest isolated coronavirus. IBV is a single-stranded positive-sense RNA virus with a genome length of about ...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12R1/93
CPCC12N7/00C12N2770/20021C12N2770/20034
Inventor 周继勇廖敏雷静李乐超莫开昆
Owner ZHEJIANG UNIV
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