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QX type chicken infectious bronchitis virus strain and application thereof

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

Active Publication Date: 2019-10-22
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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  • QX type chicken infectious bronchitis virus strain and application thereof
  • QX type chicken infectious bronchitis virus strain and application thereof
  • QX type chicken infectious bronchitis virus strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0027] In March 2018, a strain of CK / CH / LAH / 1806 virus was isolated from infected chickens in a large-scale chicken farm in Anhui. Specific method: Aseptically collect the trachea and kidney tissues of affected chickens, cut the tissues into small pieces, add 2000U penicillin and streptomycin in PBS and grind them thoroughly, homogenate to make a 30% suspension, freeze and thaw three times repeatedly, and centrifuge at 3000r / min After 15 minutes, take the supernatant, filter and sterilize, and inoculate 5 10-day-old SPF chicken embryos through the allantoic cavity route, 0.2mL / embryo, incubate at 37°C for 144 hours, collect the allantoic fluid of the chicken embryos, observe and record the death and pathological changes of the chicken embryos . Take the first-generation embryo fluid and inoculate 5 10-day-old SPF chicken embryos via the allantoic cavity route, 0.1 mL / embryo, and incubate at 37°C for 36-144 hours (take out...

Embodiment 2

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

[0030] The CK / CH / LAH / 1806 strain chicken embryo allantoic fluid was diluted 10-fold with sterile saline, and 10 -5 、10 -6 、10 -7 、10 -8For 4 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. Chicken embryos that died before 24 hours were discarded, and chicken embryos that died within 24 hours to 144 hours were taken out at any time, until 144 hours, all live embryos were taken out. 24h~144h after inoculation, dead chicken embryos and surviving chicken embryos with specific lesions such as dehydration, curling up, and small development (the weight of the inoculated fetus is more than 2g lower than that of the lightest fetus in the control) are judged as infection, and the EID is calculated. 50 . The virus content of different generations of CK / CH / LAH / 1806 strain is 10 6.17 ~10 6.83 EID 50 / 0.1mL.

Embodiment 3

[0031] Embodiment 3: CK / CH / LAH / 1806 strain purity test

[0032] According to "Veterinary Pharmacopoeia of the People's Republic of China" 2015 Edition Part Three, the CK / CH / LAH / 1806 strain chicken embryo allantoic fluid 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 there was no mycoplasma colony growth on the inoculated plate; ③ Cell method test: no cytopathic changes, no erythrocyte adsorption phenomenon caused by exogenous virus; ④ Chicken embryo method test: inoculate through the allantoic cavity (AS) and chorioallantoic membrane (CAM), and observe until 168h , fetal development was normal, CAM had no pathological changes, chicken embryo allantoic fluid HA test was negative; Inspection: Avian leukosis virus inspection (ELISA test)...

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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 application thereof. The preservation number of the strain is CGMCC No.17487. According to the QX type IBV separation strain CK / CH / LAH / 1806, the whole genome length is 27642 bp, the sequence of the strain is as shown in SEQ ID NO:1, and the strain has typical IBV genomecoding structure characteristics. The strain can cause typical IB clinical symptoms of SPF chicks which are 1-7 days old. The cross-immune protection efficacy study shows that the strain can provide immune attack protection of a QX type strain, also can provide good cross protection for attacking a Mass type strain and a TW1 type strain, and can be used as an ideal IBV vaccine candidate strain.

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] Chicken infectious bronchitis (Infection bronchitis, IB) is a highly contagious disease of chickens caused by infectious bronchitis virus (Infection bronchitis virus, IBV), which mainly affects the respiratory system, genitourinary system and digestive system of chickens Wait. It belongs to Class B infectious poultry diseases stipulated by the World Organization for Animal Health (OIE), and belongs to Class II animal diseases in the catalog of animal 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. In 1936, Beach and Hudson determined that the pathogen of the disease was a virus. In the following decades, IBV spread and became popular all over the world, causin...

Claims

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Application Information

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Patent Type & Authority Applications(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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