锦鲤疱疹病毒GY1506株的分离鉴定

Isolation and identification of koi herpesvirus GY1506

对江苏省某养殖场患病鲤鱼进行病毒分离及鉴定。现场采样发现,发病鱼体表黏液增多,眼球凹陷,背鳍和鳃部溃烂。剖检后见肝脏、肾脏、胆囊肿大,肠充血。取病鱼组织匀浆上清接种CCB细胞,14 d后可观察到细胞变圆、脱落、空泡化等典型的细胞病变。采用世界动物卫生组织(OIE)推荐的锦鲤疱疹病毒(KHV)检测引物进行PCR扩增,鳃和肠均能扩增出预期大小的特异性产物。序列分析显示,扩增序列与KHV-J毒株胸苷激酶(TK)基因核苷酸序列同源性为100%。根据TK基因序列建立系统进化树,证实该毒株为KHV亚洲型毒株,命名为KHVGY1506株。

One strain of koi herpesvirus（ KHV） was isolated and identified from diseased common carp from a fish farm in Jiangsu. The diseased cyprinoid manifested with typical clinical signs including enophthalmia（ sunken eyes） and haemorrhages on the skin and base of fins,and fin erosion. The postmortem examination revealed the intestines hyperemia and liver,kidney,cholecyst were obviously swollen. The CCB cells were inoculated with supernatant of the diseased fish tissues and 14 days later,typical cell pathological changes could be observed,such as roundout and exfoliation. Positive results were obtained from all detected gill and intestine samples according to a standard PCR diagnosis of Cy HV-3 proposed by Office International des Epizooties（ OIE）. Homology analysis of nucleotide sequences showed that the thymidine kinase（ TK） gene from infected cyprinoid shared 100% sequence identity to that of the KHV-J. Based on the TK gene sequences,phylogenetic tree was constructed and the result indicated that the viral strain in our study belongs to Asian genotype,named KHV-GY1506.