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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。
Virus isolation and identification of sick carp in a farm in Jiangsu Province. Site sampling found that the incidence of fish surface mucus increased, eye depression, dorsal fin and gill ulceration. After the autopsy, see the liver, kidney, gallbladder enlargement, intestinal congestion. Take diseased fish tissue homogenate supernatant inoculated CCB cells, 14 days after the cells can be observed round, fall off, vacuolar and other typical cytopathic effect. Using KOV detection primers recommended by the World Animal Health Organization (OIE) for PCR amplification, gill and intestine can amplify specific products of expected size. Sequence analysis showed that the nucleotide sequence homology between the amplified sequence and the KHV-J strain thymidine kinase (TK) gene was 100%. According to the sequence of TK gene, phylogenetic tree was established, which confirmed that the strain was KHV Asian strain named KHVGY1506.