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Chicken infectious bronchitis attenuated strain s1 gene and its attenuated strain and application

A technology of bronchitis and chicken infectivity, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve problems such as unexplained protective effect of bronchitis virus

Active Publication Date: 2017-05-17
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above patents all involve some new attenuated chicken infectious bronchitis vaccine strains, which can be used to prepare live vaccines, but none of these strains have shown that they can have a good protective effect on the glandular stomach type chicken infectious bronchitis virus

Method used

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  • Chicken infectious bronchitis attenuated strain s1 gene and its attenuated strain and application
  • Chicken infectious bronchitis attenuated strain s1 gene and its attenuated strain and application
  • Chicken infectious bronchitis attenuated strain s1 gene and its attenuated strain and application

Examples

Experimental program
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Effect test

Embodiment 1

[0057] Embodiment 1: the preparation of chicken infectious bronchitis virus attenuated vaccine strain (IBV) of the present invention

[0058] IBV virulent strain is chicken infectious bronchitis virus YBX strain

[0059] Infectious bronchitis virus strains are attenuated by passaging, using 9-10 day-old SPF (specific pathogen free, specific pathogen free) chicken embryos to carry out continuous passaging to attenuate the strains, and inoculate 3 chicken embryos through the allantoic cavity for each passage, continuously Passed to 170 generations, and carried out weakening evaluation test. Inoculate 1-day-old SPF chickens with F30, F65, F90, F115, F125, F145, F160 and F170 generation strains respectively, and inject 1ml into each chicken by intranasal, oral or intramuscular injection, among which F145, F160 and F170 generation virus-inoculated chicks , no morbidity and death were found, indicating that the strain has been completely attenuated in the F145, F160 and F170 genera...

Embodiment 2

[0060] Embodiment 2: the cultivation of attenuated vaccine strain chicken infectious bronchitis virus YB160 strain of the present invention and its gene stability

[0061] 1. Materials and methods

[0062] 1.1 Passage of virus strain YBX weakened

[0063] The YBX strain was attenuated by continuous passage using 9-10-day-old SPF chicken embryos. Inoculate 3 chicken embryos through the allantoic cavity for each generation of virus, inoculate 0.1ml of each embryo, incubate at 37°C for 72 hours (discard dead embryos within 24 hours), observe the chicken embryos twice a day, if the chicken embryos are within 24-72 hours If it dies, store it at 4°C. After 72 hours, all the grafted embryos were taken out and placed at 4°C for 4 to 24 hours to observe and record the lesions of the chicken embryos, and the allantoic fluid of the chicken embryos was aseptically collected for the next generation. Chicken embryos that survived 72 hours but had typical lesions and clear allantoic fluid...

Embodiment 3

[0100] Embodiment 3: vaccine preparation

[0101] 2.1 Preparation of chicken infectious bronchitis live vaccine (YB160 strain)

[0102] Take YB160 strain virus seeds (preservation number is CCTCC.V201235), dilute 100,000 times with sterilized normal saline, inoculate in the allantoic cavity of 9-10 day-old SPF chicken embryos, inoculate 0.1ml per embryo, seal the pinhole after inoculation , and incubated at 37°C. Discard the 24h dead embryos, observe the chicken embryos until 72h, put the air chamber of the 30-72h chicken embryos upward, and cool them at 4°C for 4-24h. Harvest chicken embryo allantoic fluid, mix several chicken embryo allantoic fluids into a group, put them in a sterilized bottle, store them at 2-8°C, and do a sterility test at the same time, and the virus content is 10 7.0 EID 50 / 0.1ml. The chicken embryo allantoic fluid that passed the sterility test and virus content determination was mixed with the sucrose gelatin protectant in a ratio of 1:1 (volume ...

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Abstract

The invention relates to a chicken infectious bronchitis attenuated strain S1 gene and an attenuated strain containing the attenuated strain S1 gene. The polypeptide encoded by the S1 gene of the attenuated strain can stimulate the body to produce neutralizing antibodies, and the infectious bronchitis YB160 strain containing the S1 gene of the attenuated strain can be used to prepare chicken infectious bronchitis vaccine compositions or subunit vaccines, which can effectively prevent and control Attacked by IB epidemic strains, it has better protection against glandular stomach type infectious bronchitis. The invention also relates to a diagnostic agent for chicken infectious bronchitis prepared from the attenuated strain of chicken infectious bronchitis of the invention. The present invention also relates to a chicken infectious bronchitis virulent strain S1 gene homologous to the attenuated chicken infectious bronchitis strain S1 gene of the present invention and a chicken infectious bronchitis strong virus strain containing the strong virus strain S1 gene , the virulent strain can be used for efficacy test of anti-infectious bronchitis vaccine or subunit vaccine or vaccine composition.

Description

technical field [0001] The invention belongs to the technical field of poultry biopharmaceuticals, and relates to an S1 gene of an attenuated chicken infectious bronchitis strain, an attenuated chicken infectious bronchitis strain containing the S1 gene of the attenuated strain, and an application of the attenuated strain in preparing vaccines. Background technique [0002] Avian infectious bronchitis (IB) is an acute, highly contagious viral disease of chickens. In my country, the incidence of respiratory pathological IB has been reported since 1953, and nephropathy IB was first discovered in Guangdong in 1982. In recent years, IB has been prevalent all over the country, most of which are nephropathy. Since 1995, Jiangsu, Shandong, Shanxi, Anhui and other provinces of my country have occurred a type of glandular gastropathy type IB with growth retardation, extreme emaciation, and diarrhea as the main symptoms, and glandular gastric enlargement as the main pathological featu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/50C12N7/00A61K39/215A61P31/14A61P11/00A61K49/00C12R1/93
CPCC12N7/00C12N2770/20021C12N2770/20034G01N33/56983
Inventor 张许科孙进忠白朝勇
Owner PU LIKE BIO ENG
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