鸡传染性支气管炎病毒中国地方流行株的分离与鉴定  

The Isolation and Characterization of Avian Infectious Bronchitis Virus Isolates in China

作　　者：任涛[1] 廖明[1] 罗开健 曹伟胜 辛朝安 

机构地区：[1]华南农业大学动物医学系农业部养禽与禽病重点防治实验室,广州510640

出　　处：《畜牧兽医学报》2002年第4期362-365,共4页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金项目 ( 398932 90 -2 )

摘　　要：对海南省和广西省的几个发生呼吸罗音、鸡群全群发病、死亡率不高的疑是鸡传染性支气管炎病 (IB)的鸡群 ,进行了病毒分离和鉴定。结果分离到了 4株IBV分离株 (HaN 1/95、HaN 2 /95、GX 1/98、GX 2 /98) ,并对分离病毒进行了病毒的致病性、病理组织切片、鸡胚矮小化、对NDV的干扰、电镜特征等生物特性鉴定及IBV基因组特异性 3’末端非编码区的一段保守序列的RT PCR和巢氏PCR鉴定。结果表明 4株IBV分离株 ,均能产生呼吸困难、罗音等临床症状 ;病理组织切片 ,气管、肾脏等组织表现为上皮细胞受损、固有层充血、出血和淋巴细胞的浸润及组织坏死 ;对NDV B1株有明显的干扰作用 ;其各自的传代物都有明显的致鸡胚矮小化作用 ;在透射电镜下观察到典型的冠状病毒粒子 ,多数近似为圆形 ,直径 75～ 2 0 0nm ,有囊膜 ,表面有一圈杆状纤突排列成花冠状 ,符合IBV的典型特征 ;基因组 3’末端UTR的RT PCR和巢氏PCR鉴定 ,4个分离株分别都扩增到 2 93bp和 172bp的特异目的片段 ,通过测序比较毒株间的核苷酸序列同源性在 99%以上 ,与GeneBank中IBVH5 2等标准株的 3’末端UTR的核苷酸序列高度同源。以上结果表明 4株分离病毒都是IBV ,为下一步的IBV分子流行病学的研究打下了基础。Four isolates,HaN-1/95,HaN-2/95,GX-1/98 and GX-2/98,were isolated from suspected flocks in Hainan and Guangxi province,respectively,and characterized by the experiments of pathogenicity,histopathology,dwarfing embryonated chicken eggs,interfering with the propagation of NDV-B1,electron microscopy,reverse transcription polymerase chain reaction (RT-PCR) and nested PCR.The results showed:The four isolates could result in respiratory and kidney diseases,histologically,lesions of respiratory and kidney epithelial cells,and hyperemia and hemorrhages and massive lyphoid cells infiltration and necrosis of the lamina propria;the propagation of NDV-B1 strain was seriously interfered by the 4 isolates respectively;the embryonating chicken egg passages of 4 isolates could dwarf with chicken embryos;the spherical virons 75～200 nm in diameter and surface covered with spike like corona were observed under electron microscope;the 3' non-coding ends of genome of 4 IBV isolates were amplified by reverse transcription polymerase chain reaction (RT-PCR) and nested PCR,all of four field isolates generated both 293 bp and 172 bp as expected;the nucleotide sequences similarities of the 4 isolates compared with each other were above 99%,with H52 reference strain were highly homologous respectively.On the basis of all above-mentioned results,the 4 isolates were classified as IBV.The study got a good preparedness for further study on molecular epidemiology of the 4 IBV isolates.

关 键 词：鸡 传染性支气管炎病毒 分离 鉴定 中国地方流行株 

分 类 号：S852.659.6[农业科学—基础兽医学]