羊传染性脓疱病毒的分离鉴定

Isolation and identification of Orf virus

为了对羊传染性脓疱病毒的分离株ORF027基因、ORF059部分基因进行克隆、序列分析以及构建系统进化树,本研究应用犊牛睾丸细胞从病羊痂皮病料中分离获得了1株羊传染性脓疱病毒,命名为Orf-ys,在电镜下观察到了典型的病毒粒子。应用PCR方法扩增获得了2条基因片段,PCR产物克隆至pMD18-T载体,鉴定后测序。序列分析结果表明Orf-ys的2个基因分别与参考毒株的相关基因核苷酸同源性高达97.6%～98.6%和96.9%～98.4%。基因进化树比较显示,与美国OV-IA82株和意大利OV/C2、OV/mi-90株的进化关系最近。

An Orf virus was isolated in Jilin province from crust materials of lambs with sore mouth symptom by passaging through the bovine testicle cells. The virus was identified by electron microscopy observation and PCR using Orf specific primers. The PCR products were cloned and seuqenced. The phylogenetic tree analysis indicated that there was a close genetic relationship between OrfV-ys and OV-IA82,OV/C2 strain isolated from the USA and Europe.