利用深度测序技术发现新疆库蚊黄病毒  

Identification of Culex Flavivirus by deep sequencing approach in Xinjiang, China

作　　者：刘然[1] 郑旸[2] 党荣理[1] 张桂林[1] 孙响[1] 刘晓明[1] 吴晓燕[2] 李裕昌[2] 杨银辉[2] 

机构地区：[1]新疆军区疾病预防控制中心,830011 [2]军事医学科学院微生物流行病研究所

出　　处：《中华微生物学和免疫学杂志》2014年第7期513-516,共4页Chinese Journal of Microbiology and Immunology

基　　金：自然科学基金面上项目（81171663）

摘　　要：目的：对我国新疆地区蚊虫携带的病毒类群进行调查，并快速发现医学重要的蚊媒病毒。方法利用Illumina深度测序技术筛查野生库蚊体内的病毒源性RNA，利用RT-PCR进行验证，通过序列比对分析病毒亲源关系。结果深度测序技术共检出144条库蚊黄病毒特异性读序，拼接获得7个不连续的病毒基因组重叠群片段。 RT-PCR扩增产物能够覆盖重叠群间隙序列。在此基础上，重构获得病毒基因组5′和3′末端片段（约678 nt、411 nt）。对5′末端序列比对分析结果表明，新疆库蚊黄病毒属于北美/亚洲基因亚型，与国内发现的库蚊黄病毒共同形成一个独立分支，核苷酸序列同源性为98．2％～99．5％，氨基酸序列同源性高于95．5％。结论深度测序技术成功应用于新疆地区虫媒病毒的快速甄测，首次从新疆地区发现库蚊黄病毒，为虫媒病毒监测提供了一条新途径。Objective To investigate the diversity of mosquito-borne viruses in Xinjiang , China, and to identify mosquitos-borne viruses of medical importance rapidly .Methods The virus-derived RNAs in mosquitos captured in wild were screened and confirmed by using Illumina deep sequencing approach and reverse transcription PCR , respectively .The alignment analysis was performed by using gene sequences from GenBank.Results One hundred and forty-four Culex Flavivirus （ CxFV, Flavivirus genus, Flaviviridae） specific sequences were identified .The overlapping reads were assembled into 7 uncontinuous viral genomic contigs.The gaps between the contigs were further filled by RT-PCR products, which resulted in reconstruc-tion of viral genomic 5′and 3′terminus （687 nt and 411 nt）.Phylogenetic analysis showed that the newly identified CxFV belonged to America/Asian genotype , which specifically clustered into a clade with other CxFV strains from China mainland ,sharing 98.2%-99.5%homologies in nucleotide sequences and 99.5%in amino acids sequences among them .Conclusion Illumina deep sequencing approach was successfully applied to arthropod-borne virus surveillance .The recently emerged Culex Flavivirus was detected for the first time in Xinjiang, China.

关 键 词：深度测序 库蚊黄病毒 虫媒病毒 新疆 

分 类 号：R373[医药卫生—病原生物学]