中华按蚊分离的版纳病毒全基因组序列测定与分子遗传进化分析

Genome sequencing and phylogenetic analysis of Banna virus(genus Seadornavirus, family Reoviridae) isolated from Anopheles sinensis

目的对分离自中华按蚊的版纳病毒进行病毒全基因组核苷酸序列测定和分子遗传学分析。方法使用分子病毒学方法测定病毒全基因组12节段核苷酸序列,以生物信息学软件进行序列比对、同源性和分子遗传进化分析。结果 2012年从云南省中缅边境地区中华按蚊分离的版纳病毒(YN12234病毒株)能在C6/36细胞引起细胞病变并稳定传代,该病毒为12节段RNA病毒。根据病毒VP1蛋白(RdRp)氨基酸序列进行的分子遗传进化分析发现,YN12234病毒属于呼肠孤病毒科东南亚12节段RNA病毒属版纳病毒。对第12节段基因序列的进一步分析显示,YN12234病毒属于A2基因型版纳病毒。系统进化分析结果还显示,从中华按蚊分离的版纳病毒与此前从中华按蚊分离的版纳病毒处在不同进化分支,且不同蚊虫分离的版纳病毒并非聚类在共同的进化分支。结论从中华按蚊分离的YN12234病毒株属于含有12节段的版纳病毒,分子进化分析结果提示包括云南省分离的YN12234病毒在内的目前国内外从蚊虫分离的版纳病毒并不存在病毒与蚊虫种类之间的媒介适应性。

Objective To determine the whole genome nucleotide sequence and carry out molecular genetic analysis of Banna virus (BAV)isolated from Anopheles sinensis.Methods The 12-segment nucleotide sequence of BAV was determined by molecular virological methods.Sequence alignment,homology,and molecular phylogenetic analysis were performed by using bioinformatics software.Results The BAV isolated from An.sinensis (YN12234 strain)caused cytopathic effect (CPE)and stable passage in C6/36 cells.The virus was composed of 12 segments of double-stranded RNA (dsRNA).Phylogenetic analysis of the viral VP1 protein (RdRp)amino acid sequence revealed that the YN12234 virus belongs to BAV of genus Seadornavirus of family Reoviridae.Further analysis of the 12^th segment of the gene sequence revealed that the YN 12234 virus belongs to the BAV genotype A2.The results of phylogenetic analysis also showed that the BAV isolated from An.sinensis in this study was in a different evolutionary branch from the BAV previously isolated from An.sinensis,and the BAV isolated from different mosquitoes was located in the different evolutionary cluster.Conclusion The YN12234 strain isolated from An.sinensis belongs to the BAV with 12 segments.The results of molecular evolution analysis suggest that vector adaptability between virus and mosquito species does not exist for the BAV isolated from the mosquitoes at home and abroad,including the YN12234 virus isolated in Yunnan province.