绵羊痘病毒山东流行株的分离鉴定与T4肽基因序列分析

Isolation and identification of sheep pox virus isolated from Shandong and analysis of T4 peptide gene sequence

从山东东营疑似绵羊痘病羊的肺脏组织中分离到1株病毒。取疑似绵羊痘病羊组织病料研磨、冻融、离心后分别接种11日龄SPF鸡胚绒毛尿囊膜和牛睾丸继代细胞,鸡胚接种部位出现明显的痘斑;牛睾丸细胞出现聚集、变圆等明显的细胞病变。利用1对绵羊痘病毒T4肽基因特异性引物进行了PCR扩增,获得与预期大小一致的约300 bp的片段。将所得序列与GenBank收录的绵羊痘病毒、山羊痘病毒等毒株相关序列进行比较分析。结果表明,该分离株与国外其他绵羊痘病毒株具有较高的同源性,达97.4%～99.3%;与山羊痘病毒株同源性为96.4%。通过试验初步证明所分离的病毒为绵羊痘病毒,并将该毒株命名为绵羊痘病毒DY株。

One virus strain was isolated from lung of a sheep suspected of carrying sheep pox virus, and it was named as DY strain. After rubbing, freeze-thawing and centrifuging, the tissue extracting solution was inoculated to 11 days SPF embryo and bovine testicle cell. Signif- icant pockmarks couuld be found on inoculation area of embryo, and some cytopathic effects appeared on the cells. T4 peptide gene fragments at about 300bp were amplified by PCR. The gene sequence was compared with the associated sequences from several pox virus published in GenBank. The sequence analysis showed that the viral isolate has high homology with sheep pox virus （97.4% - 99. 3% ）. The homology with goat pox virus is 96.4%. On the basis of all above mentioned results, the isolate was classified as sheep pox virus.