绵羊痘病毒固原株的分离与鉴定  

Isolation and identification of sheeppox virus Guyuan strain

作　　者：王曼[1] 叶奕优[2] 赵志荀[1] 吴国华[1] 颜新敏[1] 朱学亮[1] 李应国[3] 朱海霞[1] 李健[1] 张强[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,甘肃兰州730046 [2]深圳出入境检验检疫局动植物检验检疫技术中心,广东深圳518045 [3]重庆出入境检验检疫局,重庆400020

出　　处：《中国兽医科学》2015年第4期402-406,共5页Chinese Veterinary Science

基　　金：国家质检总局公益性行业科研专项(201310093);国家自然科学基金项目(31201892);甘肃省自然科学基金项目(1208RJZA101);国家高技术研究发展计划(863)项目(2012AA101304)

摘　　要：取发病绵羊的皮肤痘疹研磨制成病料样品悬液,将该悬液感染未免疫的健康绵羊,同时接种原代绵羊羔羊睾丸(LT)细胞,盲传5代后,转接Vero细胞,进行间接免疫荧光试验和PCR鉴定。样品悬液感染未免疫的健康绵羊后出现绵羊痘的典型发病症状,经过LT细胞传代后转接Vero细胞,出现了特征性细胞病变;间接免疫荧光试验和PCR均检测出病毒存在。PCR产物的核酸序列测定结果显示,分离毒株的RPO30基因与已发表的绵羊痘病毒毒株的相应基因的同源性在99%以上。上述结果表明,本研究从宁夏固原市疑似绵羊痘病毒感染的绵羊体内成功分离到1株绵羊痘病毒,并将其命名为绵羊痘病毒固原株。The uninfected healthy sheep were infected with the suspension made from the skin rash of sheep,and infected to primary cells of sheep lamb testicles.After 5blind passages,the infected products were harvested and infected to Vero cells.Then the virus suspension was identified by the indirect immunofluorescence assay(IFA)and PCR.The classical symptom and featured CPE were occurred in the infected sheep and Vero cells respectively.IFA and PCR results suggested that the virus particles examined in the two samples.Sequencing results showed the homology of RPO30 gene is more than 99% between the isolated virus strain in the present study and the reported strains.The above-mentioned results showed that the sheeppox virus strain,designated as GY strain,was isolated successfully from the sheep suspected sheeppox virus infection.

关 键 词：绵羊痘病毒 分离鉴定 间接免疫荧光试验 PCR 

分 类 号：S852.654[农业科学—基础兽医学]