一种新的柯萨奇病毒A组16型中国湖北分离株(CV-A16WH16)全基因组序列测定及分析  

Full genome DNA cloning and sequence analysis of a novel coxsackievirus A16 strain(CV-A16 WH16) isolated in Hubei province of China

作　　者：刘希佳[1,2] 何小华[1] 刘万红[1] 

机构地区：[1]武汉大学基础医学院,武汉430071 [2]中国科学院武汉病毒研究所,武汉430071

出　　处：《中国病毒病杂志》2015年第2期96-103,共8页Chinese Journal of Viral Diseases

基　　金：国家自然科学基金(81171577);武汉大学医学部协同创新项目(266078)

摘　　要：目的对分离自2010年中国湖北手足口患者的柯萨奇病毒A组16型(coxsackievirus A16,CVA16)病毒WH16株进行全基因组克隆、序列测定及进化分析。方法利用RT-PCR扩增覆盖基因组全长的cDNA片段,利用Over-lap PCR进行分步连接,得到CV-A16 WH16的全基因组cDNA克隆。对cDNA克隆进行测序,并利用BioEdit 4.8.8、EditSeq 5.01和MEGA 6.06软件进行序列分析和进化树构建。结果CV-A16 WH16基因组(未包含多聚腺苷酸尾)长度为7 406nt;WH16与CV-A16TS10/07、CV-A16原型株G-10以及EV71原型株BrCr的核苷酸一致性分别为97.7%、79.1%和79.5%,而氨基酸一致性分别为98.9%、94.7%和88.7%。全基因组进化树分析表明,CV-A16 WH16与CV-A16TS10/07亲缘关系最近,与G-10株存在显著差异;开放阅读框核苷酸序列进化树分析表明,CV-A16 WH16与EV71BrCr的亲缘关系较近。结论进化树分析表明,与G-10相比,CV-A16 WH16部分区域的核苷酸序列和BrCr的亲缘关系更近。Objective The full-length genome of a coxsackievirus A16（CV-A16）strain（WH16）isolated from a hand,foot,and mouse disease patient in Hubei province,China,in 2010 was cloned and its sequence characteristics were analyzed. Methods cDNA fragments covering the whole virus genome were amplified by using RT-PCR,and further linked by using over-lap extension PCR to obtain the full-length genome clone.Sequence analyses and phylogenetic analyses were performed by using BioEdit 4.8.8,EditSeq 5.01 and MEGA 6.06 software. Results The CV-A16 WH16 genome is 7 406 nt in length（excluding polyA tail）and shares 97.7%,79.1%and 79.5% nucleotide identity,and 98.9%,94.7%and 88.7%amino acid identity with the CV-A16TS10/07,CV-A16G10 and EV71BrCr,respectively.Phylogenetic analysis of the whole genome sequence suggested that WH16 has the closest relationship with CV-A16TS10/07;whilst the phylogenetic tree analysis based on sequence of open reading frame classified WH16 and EV71BrCr into one group. Conclusions The phylogenetic analysis implies that the relationship of the CV-A16 WH16 is closer to BrCr than to G-10.

关 键 词：柯萨奇病毒A16 全基因组cDNA克隆 序列分析 进化树 

分 类 号：R440[医药卫生—诊断学]