2000～2007年广西鸡传染性法氏囊病病毒的分子流行病学  

Molecular Epidemiology of Infectious Bursal Disease Virus in Guangxi during the Period of 2000 to 2007

作　　者：何秀苗[1] 韦平[2] 官丁明[1] 阳秀英[2] 秦爱建 

机构地区：[1]广西民族大学,广西南宁530006 [2]广西大学,广西南宁530004 [3]扬州大学,江苏扬州225009

出　　处：《病毒学报》2009年第6期437-444,共8页Chinese Journal of Virology

基　　金：广西自然科学基金项目(桂科自0728058);广西科技攻关项目(2007A04024;0537008-3A);扬州大学江苏省动物预防医学重点实验室开放课题(K06012)

摘　　要：应用逆转录酶-聚合酶链式反应（reverse transcriptase polymerase chain reaction,RT-PCR）技术,对2000-2007年间收集于南宁、玉林、北海和梧州4个广西主要养禽地区的临床疑似传染性法氏囊病（infectious bursal dis-ease,IBD）病鸡的法氏囊组织进行检测,检测呈阳性的病料通过9d龄鸡胚的绒毛尿囊膜（chorio-allantoic mem-brane,CAM）接种的方法进行鸡传染性法氏囊病病毒（Infectious bursal disease virus,IBDV）的分离和传代,然后设计针对病毒VP2基因高变区（vVP2）的引物对分离株进行RT-PCR扩增并对其进行核苷酸序列测定,与参考毒株的相应序列及其关键位点进行分析比较并绘制遗传进化树。结果成功分离到27株IBDV,其中BH09、BH11、TZ（3）、050222、YL051、NN0603、NN0611和QX0602等17个分离株（占62.96%）可能为超强毒（very virulent IB-DV,vvIBDV）,与其它已发表的vvIBDV毒株的序列同源性较高,在遗传进化树上可分成3个分支，与常用疫苗株的亲缘关系则较远；NN040124和YL052属中等偏强毒力株，与经典毒力株52～70和STC的亲缘关系较近；YLZF2、040131等8株属于弱毒株，与经典毒力株CU1具有较高的同源性。本研究的结果表明，近7年来在广西全区的鸡中流行的主要为vvIBDV毒株，各地毒株来源复杂，部分IBDV的抗原性可能已经发生漂变。Tissue samples of Fabricius＇ bursa collected from Nanning, Yulin, Beihai and Wuzhou in the provinces of Guangxi in China during the years of 2000-2007, were detected by a established reverse tran- scriptase polymerase chain reaction （RT-PCR） technique for IBDV. Viral isolation was performed on the positive samples by chicken embryo inoculation via chorio-allantoie membrane （CAM）. Results showed that 27 isolates of IBDV were obtained. A set of primers were designed to amplify the vVP2 of 27 isolates by RT-PCR and the PCR products were sequenced. The sequences of all the isolates and reference viruses were analyzed and compared, and their phylogenetic trees were constructed based on the nucleotide sequences. The results indicated that isolateBH11, TZ（3）, 050222, YL051, NN0603, NN0611andQX0602 etc, altogether 17 isolates, which accounted for 62.96 percent of total isolates, were identified to be very virulent IBDV （vvIBDV） and have the highest homology to vvIBDV reference strains. In the phylogenetic analysis, they are divided into 3 groups and have a long distance to commonly used vaccine stains. Isolate NN040124 and YL052 were identified as intermediate-plus virulent strains and showed a highest homology to classical strains of 52-70 and STC. 8 isolates of YLZF2, 040131 etc were identified as attenuated vaccine strains and showed a highest homology to classical strain of CU1. The results from the study demonstrated that the viruses prevailing in chickens in these 4 regions in Guangxi province in the recently 7 years were vvIBDV and their origins were complex. The antigenicity of some isolates may have been drifted.

关 键 词：RT-PCR 超强毒株 经典毒株 VP2基因高变区 分子流行病学 

分 类 号：S852.65[农业科学—基础兽医学]