S1 gene of low virulent strain of infectious bronchitis and low virulent strain and application thereof

A bronchitis, chicken infectious technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problem of not showing the protective effect of bronchitis virus and so on

Active Publication Date: 2014-06-11
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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, bu

Method used

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  • S1 gene of low virulent strain of infectious bronchitis and low virulent strain and application thereof
  • S1 gene of low virulent strain of infectious bronchitis and low virulent strain and application thereof
  • S1 gene of low virulent strain of infectious bronchitis and low virulent strain and application thereof

Examples

Experimental program
Comparison scheme
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 an S1 gene of a low virulent strain of infectious bronchitis and the low virulent strain comprising the S1 gene of the low virulent strain. Polypeptide coded by the S1 gene of the low virulent strain can stimulate a body to generate a neutralizing antibody, and a YB 160 strain of infectious bronchitis, comprising the S1 gene of the low virulent strain, can be used for preparing a vaccine composition or a subunit vaccine for treating infectious bronchitis, can effectively prevent attack from the IB prevalent strain and has a good protective effect on glandular stomach infectious bronchitis. The invention further relates to an infectious bronchitis diagnostic reagent prepared from the low virulent strain of infectious bronchitis. In addition, the invention further relates to the S1 gene of a virulent strain of infectious bronchitis homologous with the S1 gene of the low virulent strain of infectious bronchitis and the virulent strain of infectious bronchitis comprising the S1 gene of the virulent strain. The virulent strain can be used for inspecting potency of a vaccine or subunit vaccine or a vaccine composition for treating infectious bronchitis.

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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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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