2009年云南省柯萨奇病毒B3型分离株A103／KM／09全基因序列分析  

Complete nucleotide sequence of a human coxsackievirus B3 strain A103/KM/09 isolated in Yunnan province, 2009

作　　者：杨卉娟[1] 刘建生[1] 张云昆[1] 马忠飞[1] 赵卫中[1] 潘玥[1] 陈俊英[1] 邵聪文[1] 马绍辉[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所云南省重大传染病疫苗研发重点实验室,昆明650118

出　　处：《中华流行病学杂志》2013年第12期1208-1212,共5页Chinese Journal of Epidemiology

基　　金：云南省应用基础研究项目（2013FZ136）

摘　　要：目的分析2009年云南省脑炎患儿粪便标本分离的柯萨奇病毒B3型分离株A103／KM／09全基因序列，了解其遗传特性。方法设计针对柯萨奇病毒B3型引物，提取病毒RNA、RT-PCR扩增和产物直接测序获得全基因组序列，利用Geneious和Mega5．05软件进行核苷酸、氨基酸序列比对及其系统进化分析，应用RDP3和SimPlot3．5．1重组软件对序列进行重组分析。结果该分离株全基因组核苷酸序列长度为7389bp。其核苷酸和氨基酸序列与其原型株Nancy的同源性分别为81．0％和95．7％；与其他柯萨奇病毒B3型毒株的各区段核苷酸和氨基酸同源性分别为81．0％～88．0％和95．7％～98．0％；进化分析发现CVB3可分为5个分支（I～V），核苷酸之间的差异为16．2％-24．3％；中国分离株属V分支，又可分为A～D亚分支，核苷酸之间的差异为4．3％～11．4％。并发现A103／KM／09株基因序列在非结构区可能存在重组事件。结论柯萨奇病毒B3型分为5个基因型，云南分离株A103／KM／09属于V—D亚型，而中国分离株已经发生一定进化。Objective To analyze the genetic characterization of the complete genome from a human coxsackievims B3 strain A103/KM/09 isolated in Yunnan province, 2009. Methods By using RT-PCR, all the eight fragments which containing about 1000 nucleotides and covering full viral genome, were sequenced. By using Mega 5.05, Geneious, RDP 3 and SimPlot 3.5.1 software, sequences were aligned with other enterovirus reference sequences. Phylogenetic and recombination analysis were also carried out. Results The A103/KM/09 isolate genome showed 7389 nucleotides in length, encoding for 2185 amino acids. In the complete genome, the homology of nucleotide and amino acid among the seven coxsackievirus B3 isolates were 81.0%-88.0% and 95.7%-98.0%, respectively. There appeared 81.0% and 95.7% homology when compared with that of Nancy prototype strain. Results from the Phylogenetic analysis showed that the coxsackievirus B3 formed five distinct clades, I - V. Nucleotide divergence rates between clades were 16.2%-24.3%. The A103/KM/09 strain belonged to clade V. Clade V was further divided into four sub-clades, A-D. The nucleotide divergence between sub-clades was 4.3%-11.4%. Putative recombinant event for A103/ KM/09 was detected. Conclusion All coxsackievirus B3 isolates could be divided into five clades, with A103/KM/09 strain belonged to Clade V -D. Evolution of coxsackievirus B3 had occurred in China.

关 键 词：柯萨奇病毒B3型 全基因序列 序列分析 

分 类 号：R725.1[医药卫生—儿科]