福建省首株黄热病毒株的分离鉴定和全基因组序列分析

Isolation, identification and genomic sequence analysis of the first yellow fever virus strain in Fujian province

目的从福建省2016年输入性黄热病病例标本中分离黄热病毒(YFV)并分析其生物学特征。方法将16份黄热病毒核酸阳性血清、尿液、唾液样品分别接种C6/36细胞,YFV实时荧光RT-PCR法鉴定检测培养物中特异性病毒核酸,通过高通量测序获得病毒全基因序列并绘制系统进化树。结果从1例患者发病后3天的血清样品分离到一株病毒,通过YFV实时荧光RT-PCR鉴定为YFV病毒;BLAST分析显示该毒株全基因序列与2016年我国首例输入性黄热病病例中分离得到的毒株CNYF01R/2016(KX268355)序列完全一致;系统进化树显示该毒株与1971年安哥拉流行株Angola71分属同一进化树分支,与疫苗株17D vaccine strain(X03700)存在明显的差异,表明本研究分离到的YFV毒株是属于安哥拉型YFV野毒株。结论福建省成功从2016年输入性黄热病病例样品分离到一株安哥拉型YFV毒株。

Objective To isolate and identify the yellow fever virus(YFV)from the specimens of the imported yellow fever(YF)cases in Fujian province in 2016.Methods Sixteen positive YFV nucleic acid samples including serum,urine and saliva were inoculated into C6/36 cells,respectively.The isolated strain was identified by YFV real-time RT-PCR.The complete gene sequence of this strain was obtained by high-throughput next-generation sequencing,and the phylogenetic tree was drawn.Results Only one strain was isolated from the serum of one case three days after onset,and identified as a YFV strain by real-time RT-PCR.BLAST analysis showed that the complete gene sequence of this strain was identical to the strain CNYF01R/2016(KX268355)isolated from the first YF imported case in China in 2016.The phylogenetic tree showed that this strain belonged to the same phylogenetic branch as the epidemic strain Angola71 in Angola in 1971,and was significantly different from the 17D vaccine strain(X03700),indicating that the YFV strain isolated in this study belonged to the wild YFV strain of Angola genotype.Conclusions An Angola genotype YFV strain was successfully isolated from samples of imported YF cases in Fujian Province in 2016.