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A qx type chicken infectious bronchitis virus strain and its application

A bronchitis, chicken infectious technology, applied in the field of virology, can solve the problems of biological characteristics, complex and variable pathogenicity and tissue tropism, cross-protection and poor serum neutralization efficiency.

Active Publication Date: 2022-07-22
JIANGSU AGRI ANIMAL HUSBANDRY VOCATIONAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that the biological characteristics, pathogenicity and tissue tropism of different types of IBV strains are complex and changeable. According to the differences in immunogenicity, IBV can be divided into multiple serotypes, and the cross-protection between different serotypes and relatively poor serum neutralization potency

Method used

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  • A qx type chicken infectious bronchitis virus strain and its application
  • A qx type chicken infectious bronchitis virus strain and its application
  • A qx type chicken infectious bronchitis virus strain and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: CK / CH / LAH / 1806 strain isolation

[0027] In March 2018, a strain of CK / CH / LAH / 1806 virus was isolated from a diseased flock of a large-scale chicken farm in Anhui. Specific method: aseptically collected trachea and kidney tissue of diseased chickens, cut the tissue into pieces, add PBS containing 2000U penicillin and streptomycin to fully grind, homogenize to make 30% suspension, freeze and thaw three times, and centrifuge at 3000r / min For 15 minutes, the supernatant was filtered and sterilized, and then 5 SPF chicken embryos of 10 days old were inoculated through the allantoic cavity route, 0.2 mL / embryo, incubated at 37°C for 144 hours, the chicken embryo allantoic fluid was collected, and the death and lesions of the chicken embryos were observed and recorded. . Take the first generation of embryo fluid to inoculate 5 10-day-old SPF chicken embryos through the allantoic cavity route, 0.1 mL / embryo, and incubate at 37°C for 36-144 h (specifically dead embr...

Embodiment 2

[0029] Example 2: Determination of virus content of CK / CH / LAH / 1806 strain

[0030] The chicken embryo allantoic fluid of the CK / CH / LAH / 1806 strain was serially diluted 10 times with sterile normal saline, and 10 -5 , 10 -6 , 10 -7 , 10 -84 dilutions, inoculate 5 10-day-old SPF chicken embryos via the allantoic cavity route, 0.1 mL per embryo, and continue to incubate at 36°C to 37°C. The chicken embryos that died before 24h were discarded, and the chicken embryos that died within 24h to 144h were taken out at any time, and all live embryos were taken out at 144h. 24h to 144h after inoculation, the dead chicken embryos and the surviving chicken embryos showed specific lesions such as dehydration, curling up, and small development (the weight of the inoculated fetus was more than 2g lower than that of the lightest fetus in the control), which was judged as infection, and the EID was calculated. 50 . The virus content of different generations of CK / CH / LAH / 1806 strain is 10 ...

Embodiment 3

[0031] Example 3: Purity test of CK / CH / LAH / 1806 strain

[0032] According to "Veterinary Pharmacopoeia of the People's Republic of China" 2015 edition three parts, the chicken embryo allantoic fluid of CK / CH / LAH / 1806 strain was tested for sterility, mycoplasma and exogenous virus. ① Sterility test: no bacterial growth was found in the TG / TSB test tubes inoculated with chicken embryo allantoic fluid; ② Mycoplasma test: the color of the mycoplasma culture medium inoculated with allantoic fluid did not change, and no mycoplasma colony growth appeared on the inoculated plate; ③Cell method test: no cytopathic changes, and no erythrocyte adsorption caused by exogenous virus; ④Chicken embryo method test: inoculation through the allantoic cavity (AS) and chorioallantoic membrane (CAM) route, and observed for 168 hours , the fetus develops normally, the CAM has no lesions, and the chicken embryo allantoic fluid HA test is negative; Test: Avian leukemia virus test (ELISA test) was nega...

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Abstract

The invention relates to the technical field of virology, and discloses a QX-type chicken infectious bronchitis virus strain and an application thereof. The preservation number of the strain is CGMCC No.17487. The present invention provides a QX type IBV isolated strain CK / CH / LAH / 1806, the whole genome length of which is 27642 bp, the sequence is shown in SEQ ID NO: 1, and has typical IBV genome coding structure characteristics. This strain can cause typical clinical symptoms of IB in SPF chicks aged 1-7 days. Cross-immunity protection efficacy studies have shown that this strain can not only provide immune challenge protection against QX strains, but also provide good cross protection against challenge with Mass and TW1 strains, making it an ideal IBV vaccine candidate. strains.

Description

technical field [0001] The invention relates to the technical field of virology, in particular to a QX-type chicken infectious bronchitis virus strain and application thereof. Background technique [0002] Infection bronchitis (IB) is a highly contagious disease of chickens caused by Infection bronchitis virus (IBV), which mainly affects the respiratory system, urogenital system and digestive system of chickens. Wait. It belongs to the category B infectious poultry disease stipulated by the World Organization for Animal Health (OIE), and it belongs to the second category of animal epidemic diseases in the catalogue of animal epidemic diseases issued by the Ministry of Agriculture of my country. In the 1930s, Schalk and Hawn first discovered the disease from North Dakota in the United States, and in 1936 Beach and Hudson identified the disease as a virus. In the following decades, IBV was widely spread and popular around the world, causing serious economic losses to the wor...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/215A61P31/14C12R1/93
CPCC12N7/00A61K39/12A61P31/14C12N2770/20021C12N2770/20034A61K2039/525A61K2039/552
Inventor 穆晓惠陈启稳曹春光朱善元秦爱建肖文华洪伟鸣
Owner JIANGSU AGRI ANIMAL HUSBANDRY VOCATIONAL COLLEGE
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