羊口疮病毒的分离与鉴定  

Isolation and Identification of Orf Virus

作　　者：庞方圆 高日明 李旭东[1] 闫聪 李浩[1] 王艳杰 张七斤[1,3] 

机构地区：[1]内蒙古农业大学兽医学院,呼和浩特010018 [2]金宇保灵生物药品有限公司,呼和浩特010030 [3]兽用疫苗国家工程实验室,呼和浩特010010

出　　处：《中国畜牧兽医》2015年第2期467-471,共5页China Animal Husbandry & Veterinary Medicine

基　　金：兽用疫苗国家工程实验室项目

摘　　要：本试验利用犊牛睾丸细胞、羔羊睾丸细胞、MDBK、BHK-21细胞对采自内蒙古赤峰的疑似羊口疮发病羊群的唇部痂皮组织进行接种传代,分离出1株病毒,通过透射电镜负染观察和PCR检测对该分离株进行鉴定。参照GenBank中羊口疮病毒(Orf virus,ORFV)ORF059(F1L)基因的核苷酸序列,设计并合成1对特异性引物,成功地对分离株的F1L基因进行克隆、测序,并与多株参考毒株进行了同源性比对分析,结果显示经透射电镜负染观察到典型的副痘病毒粒子,与参考毒株序列相比均具有较高的同源性,均为96%以上,结果表明该分离株为ORFV,命名为OV/nm-hd。One strain of virus was isolated by passage culture in bovine testis cell,lamb testis cell, MDBK cell and BHK-21 cell of lip crust tissue collected from suspected Orf sheep in Chifeng,Inner Mongolia. The isolated virus was identified through negative staining by TEM observation and PCR detection method. One pair of specific primers was designed and synthesized referring to nu- cleotide sequence of Orf virus （ORFV） ORF059 （F1L） gene in GenBank. F1L gene of isolated vi- rus was successfully cloned and sequenced, the homology was analyzed with many reference strains. The result showed that typical brick-shaped virion was found by TEM observation, and the isolated strains had high homologies with reference strains, the homologies were all more than 96% ,illustrating that the isolated strains was ORFV,named as OV/nm-hd.

关 键 词：羊口疮病毒 分离鉴定 F1L基因 序列分析 

分 类 号：S852.65[农业科学—基础兽医学]