传染性法氏囊病毒安徽超强毒株的分离及分子鉴定  

Isolation and Molecular Identification of Very Virulent Infectious Bursal Disease Virus

作　　者：王桂军[1] 吴忆春[1] 魏建忠[1] 李郁[1] 何长生[2] 韦平[3] 

机构地区：[1]安徽农业大学动物科技学院,安徽合肥230036 [2]安徽省兽医工作站,安徽合肥230022 [3]广西大学养禽与禽病研究所,广西南宁530005

出　　处：《中国农学通报》2006年第10期17-21,共5页Chinese Agricultural Science Bulletin

基　　金：安徽省教育厅自然科学基金项目"安徽省鸡传染性法氏囊病的分子流行病学研究"(2004kj152);教育部科研重点项目"我国部分省区鸡免疫抑制病的流行病学研究"(02116)

摘　　要：应用鸡胚绒毛尿囊膜(CAM)接种﹑琼脂扩散试验、电镜观察,从临床病鸡的法氏囊组织分别分离到3株传染性法氏囊病病毒(IBDV)CH03、JG04和QJ04株,对4周龄未免疫鸡的攻毒致死率分别为92%、83%、67%,对鸡胚的半数致死量(ELD50)分别为10-6.8/0.2ml、10-5.4/0.2ml、10-4.6/0.2ml;应用逆转录酶-聚合酶链式反应(RT-PCR)扩增的IBDVVP2基因高变区(vVP2)及其序列分析的结果表明,序列共有492个核苷酸,编码164个氨基酸。关键位点的氨基酸变化特征:第一亲水区P222A、疏水区的K249Q和S254G、N279D和T284A,与超强毒参考株UK661﹑HK46﹑OKYM和CH1-97极为相似,而与致弱株Cu-1)及变异毒株Var-E存在明显差异。研究结果表明,获得的3株IBDV分离株均属超强毒株。3 strains of infectious bursal disease virus （IBDV） designated as CH03, JG04 and QJ04, were isolated and identified from bursa tissues of infected chickens in Anhui province by chicken embryo inoculation, AGP test and electronic microscopy. 4-week-old chickens, which were unvaccinated of IBDV, were inoculated with the tree isolates and experienced severe clinical disease with the mortalities of 92%, 83% and 67%, and the ELD50 of 3 isolates were 10^-6.8/0.2ml, 10^-5.4/0.2ml and 10^-4.6/0.2ml, respectively. The VP2 hypervariable regions （vVP2） were amplified by RT-PCR （reverse transcriptase polymerase chain reaction）/Nested PCR and sequence analysis of the amplified products showed that the corresponding drifts of amino acids were found at five different epitopes： P222A located in the first hydrophilic region, K249Q, S254G, N279D and T284A located in the hydrophobic region, respectively. The results demonstrated that these 3 isolates were classified to be very virulent infectious bursal disease virus （vvIBDV） since they possess the epitope characteristics of vvIBDV but no that of attenuated or variant IBDV.

关 键 词：IBDV 超强毒株 分子鉴定 

分 类 号：S852.659.4[农业科学—基础兽医学]