4株IBDV的分离鉴定及VP2基因序列分析  

Isolation and identification of four infectious bursal disease virus strains and sequence analysis of VP2 gene

作　　者：王帅涛[1] 常洪涛[1] 李欢[1] 李伟豪[1] 燕贺[1] 王川庆[1] 姚惠霞[1] 刘红英[1] 王新卫[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002

出　　处：《扬州大学学报（农业与生命科学版）》2013年第3期5-9,共5页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家农业科技成果转化资金资助项目(2011GB2D000007)

摘　　要：通过接种鸡胚绒毛尿囊膜(CAM)、DF-1细胞和琼脂扩散试验,对2010～2012年间收集于河南地区疑似鸡传染性法氏囊病病料,进行鸡传染性法氏囊病病毒(IBDV)的分离、鉴定,得到4株IBDV;应用RT-PCR对分离株VP2基因进行扩增、克隆和序列测定后,分析比较4个分离株与参考毒株的VP2序列。结果表明:LY株属于超强毒株,与已发表的vvIBDV D6948株、OKYM株、UK661株、GZ/96株亲缘关系较近,核苷酸同源性达97.9%～98.2%;ZZ株属于强毒株,与标准强毒株52/70株亲缘关系较近,核苷酸同源性为97.8%;AY、XC株不仅具有弱毒株的特征,同时又具有变异毒株的特征,属于变异弱毒株,与标准弱毒株CU1的亲缘关系最近,核苷酸同源性达99.0%～99.3%。证明河南区域同时存在着不同毒力的IBDV毒株,病毒变异呈多态性。Tissue samples of fabricius bursal were collected from Henan Province during the years of 2010-2012. 4 isolates of IBDV were obtained by chicken embryo inoculation via chorio-allantoic membrane （CAM）, cell culture with DF-1 and by the agar diffusion test. The VP2 genes of 4 isolates were amplified by RT-PCR and sequenced respectively. The results indicated that isolate LY was classified to be very virulent IBDV, and had close phylogenetic relationship with isolate D6948, isolate OKYM, isolate UK661 and GZ/96, and their nucleotide homology was 97.9%-98.2%. Isolate ZZ was classified to be classical virulent IBDV, and had close phylogenetic relationship with isolate 52/70, and the two strains nucleotide homology was 97.8%. While isolate AY and XC were identified as avirulent IBI）V and had close phylogenetic relationship with classical avirulent strain CU1, at the same time displayed some characters of the variant strain, and their nucleotide homology was 99.0%- 99.3%. The results of the study demonstrated that there were infectious bursal disease virus strains of different pathotypes and viral variation showed polymorphisms in Henan during 2010- 2012.

关 键 词：传染性法氏囊病病毒 超强毒株 VP2基因 序列分析 

分 类 号：S852.659.4[农业科学—基础兽医学] Q28[农业科学—兽医学]