云南省三带喙库蚊中分离到Toti病毒

目的对云南省蚊虫病毒分离物进行分子生物学鉴定,以明确其分类地位。方法采用C6/36细胞对病毒分离物ML-13做病毒扩增,使用多种虫媒病毒基因扩增引物进行病毒基因扩增和基因序列测定。采用推导的氨基酸序列用Mega5.0软件构建种系发生树。结果 ML-13病毒可以在蚊虫细胞增殖,用OMRV和ToV-TJ病毒特异引物扩增出目的产物,序列分析显示,ML-13病毒与OMRV-AK4、ToV-TJ、OMRV-Y61、DTV、AsTV病毒衣壳蛋白和RNA依赖RNA聚合酶基因株相似性最高,分别为76%、75%、72%、67%和67%;与Toti病毒科中的DTV、ToV-TJ病毒和OMRV处于同一进化分支。结论云南省三带喙库蚊中分离的ML-13病毒为Toti病毒,这是我国首次从三带喙库蚊中分离到Toti病毒。

Isolation of Totivirus from Culex tritaeniorhynchus in Yunnan province,China

Objective To identify the isolated virus(ML-13) from mosquitoes in Yunnan province,China by molecular biology techniques.Methods The new virus isolate(ML-13) was amplified by C6/36 cells.The viral gene was amplified and sequenced using several sets of arbovirus primers.A phylogenetic tree was constructed using Mega 5.0 based on deduced amino acid sequences.Results ML-13 virus could be amplified in C6/36 cells.Target products were obtained by amplification using specific primers for Omono River virus(OMRV) and Tianjin Totivirus(ToV-TJ).ML-13 virus showed the highest sequence similarities(76%,75%,72%,67%,and 67%) with OMRV-AK4,ToV-TJ,OMRV-Y61,DTV,and AsTV with regard to capsid protein(CP) and RNA dependent RNA polymerase(RdRp).The phylogenetic analysis showed that ML-13 was in the same branch as OMRV,ToV-TJ,and DTV in Totiviridae family.Conclusion ML-13 strain isolated from Culex tritaeniorhynchus in Yunnan province,China belongs to the Totiviridae family and is the first isolate of Totivirus from Cx.tritaeniorhynchus in China.