海南省三亚地区蚊虫细胞融合病毒鉴定与分析

Identification and analysis of Aedes flavivirus in Sanya City of Hainan Province

目的了解海南省三亚地区蚊虫携带细胞融合病毒现况。方法在2018-2019年蚊虫活动高峰期采集三亚地区蚊虫,前期运用高通量测序的方法获得了1份细胞融合病毒核酸序列,本研究根据已测序结果设计特异引物,通过RT-PCR扩增伊蚊细胞融合病毒目的片段及测序,检测蚊虫携带的细胞融合病毒阳性率,运用MEGA 7.0软件对所得病毒基因核酸序列与Genbank中已知细胞融合病毒参考序列进行遗传进化分析。结果共采集290只蚊虫,其中从224只白纹伊蚊中仅扩增得到1份细胞融合病毒(CFAV-NS3-SY1),致倦库蚊(26只)和三带喙库蚊(40只)均未检测到该病毒,蚊媒活动高峰期蚊虫带毒率为0.34%。该病毒NS3基因系统发育分析发现CFAV-NS3-SY1与分离自法国马赛的病毒株(AF411835)遗传关系最近,相似度为99.84%,因此可以将这株病毒界定为白纹伊蚊细胞融合病毒。结论三亚地区白纹伊蚊携带细胞融合病毒,应增加样本量持续开展蚊虫携带病原体监测。

Objective To investigate the cell fusing agent virus(CFAV)carried by Aedes albopictus in Sanya City.Methods Mosquitoes were collected at the peak of mosquito activity in 2018-2019.In the early stage,a copy of nucleic acid sequence of CFAV was obtained by high-throughput sequencing.Therefore,in this study,specific primers were designed according to the sequenced results,and the target segments of Aedes flavivirus were amplified and sequenced by RT-PCR.The virus infection rate of mosquitoes were tested.Mega 7.0 software was used to construct phylogenetic tree between the nucleic acid sequence of the obtained virus gene and the known reference strain of CFAVs in GenBank,and to analyze the genetic evolution relationship.Results Only one copy of CFAV was obtained from 224 Aedes albopictus,and the virus was not detected from 26 Culex pipiens quinquefasciatus and 40 Culex tritaeniorhynchus.The mosquito carrying virus rate was 0.34% at the peak of mosquito activity.The phylogenetic analysis of NS3 gene of the virus showed that CFAV-NS3-SY1 had the closest genetic relationship with the virus strain(AF411835) isolated from Marseille,France,with a similarity of 99.84%.Therefore,this virus could be defined as cell fusing agent virus.Conclusion Aedes albopictus in Sanya area carries cell fusing agent virus.It is necessary to increase the sample size and carry out continuous surveillance of mosquito borne pathogens.