1株传染性法氏囊病病毒的分离鉴定及VP2基因序列分析

doi:10.16431/j.cnki.1671-7236.2022.03.022

Abstract

Abstract: 【Objective】 The purpose of this study was to study the virulence characteristics of a strain of Infectious bursal disease virus (IBDV) isolated from Guangxi and its relationship with the sequence characteristics of VP2 gene, so as to provide reference for the epidemiological research and epidemic prevention and control of IBDV.【Method】 The virus was isolated and identified by PCR, chicken embryo subculture, adaptive culture of DF-1 cells and agar diffusion test, and its VP2 gene was amplified.The nucleotide and amino acid sequences were compared with other IBDV strains with different virulence by Mega 7.0 and MegAlign softwares, and the pathogenicity to SPF chicken was tested.The changes of IBDV copy number were determined by Real-time PCR.【Result】 One strain of IBDV was successfully isolated, and named as GX20210126 strain.SPF chicken embryos inoculated with this strain could cause bleeding, dwarfing, liver yellowing, bleeding and acitoid necrosis.Moreover, GX20210126 had good cell adaptability to DF-1 and could cause significant cytopathic changes.The agar diffusion test showed that the strain had good antigenicity, and a white precipitation line could be formed between the virus liquid and the IBDV-positive serum.VP2 gene evolution analysis showed that the GX20210126 strain was on the same branch with the international standard super virulent strain, and the amino acid similarity was between 86.8% and 99.6%.It had the highest similarity with the UK661 strain, and the amino acid similarity was 99.6%.Only two amino acid positions changed, which were D279N and I272T.The 8-day-old SPF chickens were inoculated with EID₅₀ 10^-3.8/0.1 mL, 0.5 mL per chicken.After inoculation, the chickens showed fluffiness, droopiness, white and green water feces, and other clinical symptoms.Necropsy showed muscle and liver hemorrhage, kidney urate deposition, and bursa atrophy.What was more, IBDV copy number peaked on the 5th day.【Conclusion】 The IBDV epidemic strain GX20210126 in Guangxi had the genetic sequence characteristics of a super virulent strain.Artificial infection of chickens could lead to typical clinical symptoms and pathological changes of IBDV.

Key words: Infectious bursal disease virus (IBDV); VP2 protein; isolation and identification; sequence analysis

CLC Number:

S852.65

HE Xianming, REN Guangcai, YE Junxian, LAN Hong, LIU Yufu, XIONG Ting, YANG Zekun, XU Ting, CHEN Ruiai. Isolation and Identification of a Strain of Infectious Bursal Disease Virus and Sequence Analysis of VP2 Gene[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1005-1014.

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Isolation and Identification of a Strain of Infectious Bursal Disease Virus and Sequence Analysis of VP2 Gene

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