新疆艾比湖布尼亚病毒的分离与分子生物学鉴定

Isolation and molecular characterization on Abbey Lake Orthobunyavirus （Bunyaviridae） in Xinjiang, China

目的 发现新疆艾比湖地区潜在流行的蚊媒病毒.方法 采用接种敏感宿主细胞法分离培养病毒;利用反转录聚合酶链和序列聚类分析对病毒株种属进行分子生物学鉴定.结果 从当地优势蚊种凶小库蚊（36.6％）中分离获得一株引起BHK-21细胞病变的病毒,分子生物学鉴定表明该病毒属于布尼亚科正布尼亚属（Orthobunyavirus,Bunyaviridae）成员,暂定名为艾比湖布尼亚病毒.RT-PCR扩增进一步获得长为651 bp和980 bp的病毒基因组S、M节段的部分序列,对上述序列的比对分析显示艾比湖布尼亚病毒与分离自南非的Germiston布尼亚病毒同源关系最为接近,S节段核苷酸和氨基酸序列似然率分别为90.6％和95.0％.M节段变异性高,核苷酸和氨基酸序列似然率仅为78.6％和86.1％.结论 艾比湖布尼亚病毒是国内新发现的一株蚊媒布尼亚病毒.

Objective To monitor and discover medically important mosquito-borne viruses circulating in Xinjiang,China.Methods Mosquitoes were collected from Abbey Lake wetland in Bortala,in Northern Xinjiang.Viral isolates were obtained through innoculating and serial passaging into susceptible mammalian host cells （BHK-21）,identified by cytopathogenic effect （CPE） observation and plague forming assay.Genetic identification of viral isolates was conducted by RT-PCR,sequencing and phylogenetic analysis.Results A virus strain which causing CPE on BHK-21 cells,was isolated from the predominant Culex modestus （36.6%） and tentatively designated as Abbey Lake virus.Information on molecular identification revealed that Abbey Lake virus belonged to Orthobunyavirus genus within Bunyaviridae.Partial sequences （651 bp and 980 bp） of viral genomic S and M segment showed that Abbey Lake virus was phylogenetically related to Germiston virus that uniquely found in South Africa with 90.6% nucleotides and 95.0% amino acids similarities in S segment.However,viral M segment displayed much variability with 78.6% nucleotides and 86.1% amino acid similarities,suggesting a new member of Orthobunyavirus genus was discovered in the area.Conclusion In this study,Abbey Lake virus was isolated and characterized indicating its potential circulation nature of this newly-emerged mosquito-borne virus.