新疆南疆牛冠状病毒BCV-Aks-01株分离及基因型鉴定

Isolation and Genotype Identification of Bovine Coronavirus BCV-Aks-01 Strain in Southern Xinjiang

将疑似感染牛冠状病毒(Bovine coronavirus,BCoV)病死犊牛的粪便样品,滤菌处理后接种于HCT-8细胞,盲传数代,出现细胞病变后收获细胞液。提取病毒RNA,依据NCBI登录的牛冠状病毒高度保守N基因设计1对特异性引物,对N基因进行RT-PCR扩增、测序及序列分析。结果显示,BCV-Aks-01毒株与牛冠状病毒参考株N基因核苷酸同源性为97.5%~98.6%,同BCoV参考毒株Mebus同源性为97.8%,N基因遗传进化树表明,BCV-Aks-01毒株与BCoV参考株亲缘关系较近,处于遗传进化树中同一分支上,属于冠状病毒Betacoronavirus,证实该分离株为牛冠状病毒。

The filtrating fecal Suspension from a suspected infected Bovine coronavirus calf was inoculated into HCT-8 cells. Af- ter blind passages, virus solution was harvested when the CPE appeared. RNA sample was extracted and detected by RT-PCR with special primers. The amplified fragments were sequenced. Analysis of the nucleotide sequences and construction of phylogenetic tree were based on N gene of coronavirus with computer programs. The sequencing results showed that the N gene sequence of BCV-Aks- 01 strain had 95% similarity to the reference strain from GenBank.. The genotyping of BCV-Aks-01 strain belonged to Betacoronavirus of Bovine coronavirus. The result showed that BCV-Aks-01 strain belonged to Bovine coronavirus.