羊口疮病毒威海株的分离鉴定

Isolation and Identification of Orf Virus Weihai Strain

利用犊牛睾丸原代细胞对山东省威海市某奶山羊养殖场疑似羊口疮(Orf)山羊病料进行病毒分离,通过细胞病变观察、病毒滴度和聚合酶链反应(PCR)等方法对分离毒株进行鉴定。结果表明,研磨处理的病毒液接种于犊牛睾丸原代细胞后盲传至第5代出现病变,测得第6代分离株病毒滴度为106.5 TCID50/mL。扩增羊口疮病毒特异性B2L基因,发现该毒株与34个羊口疮毒株核苷酸序列同源性高达99%,包括福建株(KC588399.1、KC568398.1)。本研究成功分离鉴定了羊口疮病毒山东威海株。

The virus strain was isolated from crust materials of lambs with Orf like-clinical symptoms from a goat farm in Weihai city of Shandong province by passage in primary bovine testicular cells （PBTC）.The virus was identified as Orf virus using cell morphology of lesions,TCID50 test and PCR method.The re-sults showed that PBTC cultures infected with crust materials displayed cytopathic effects at the fifth pas-sages.The titer of the sixth passage virus was 106.5 TCID50/mL.The specific Orf virus B2L gene was am-plified.The product was sequenced,and the isolated strain was closest to similarity of nucleotide sequence （99%）with those of Fujian strain （KC588399.1,KC568398.1）.It was concluded that the Orf virus strain was successfully isolated and identified.