2009年云南省昆明地区柯萨奇病毒B5的遗传特性分析

目的对2009年中国云南省昆明地区柯萨奇病毒B5(Coxsackievirus group B5,CVB5)的部分VP1基因序列进行分析,以了解CVB5的遗传特性。方法收集2009年中国云南省昆明地区临床诊断为无菌性脑炎患儿的200份粪便标本,设计针对CVB的通用引物,利用半巢式RT-PCR扩增部分VP1基因。PCR产物直接测序后,采用Omiga和Mega 5.05等生物学软件进行核苷酸、氨基酸序列及系统进化分析。结果共分离出7株CVB5,其部分VP1基因之间的核苷酸和氨基酸序列同源性均较高,分别为92.7%～98.5%和93.6%～100.0%;与中国其他不同地区参考株的核苷酸序列同源性为80.1%～99.1%,其中与中国山东株98388-SD-CHN-1998同源性较低;与中国其他不同地区参考株的氨基酸序列同源性为84.4%～100.0%,其中与中国河南株CoxB5-Henan-2010和中国山东株98388-SD-CHN-1998之间同源性最低;与其他不同国家参考株比较,其核苷酸和氨基酸序列同源性分别在77.5%～89.1%和92.1%～96.3%之间;与原型株Faulkner的核苷酸和氨基酸序列同源性分别为79.6%～80.9%和92.1%～95.4%。基因系统进化分析显示,分离的7株CVB5属于相对独立的1个分枝,中国流行株除98388-SD-CHN-1998外,构成1个分枝。结论 2009年中国云南省昆明地区CVB5流行株属于相同基因型。

Genetic characters of coxsackievirus B5 strains isolated in Kunming Region, Yunan Province, China in 2009

Objective To analyze the sequences of partial VP1 genes of seven coxsackievirus B5(CVB5)strains isolated in 2009 in Kunming Region,Yunnan Province,China so as to investigate the genetic characters of CVB5.Methods A total of 200 fecal specimens were collected from the children diagnosed as patients with aseptic meningitis in Kunming Region in 2009,from which partial VP1 genes were amplified by semi-nested RT-PCR using designed general primers.The PCR products were sequenced directly,of which the nucleotide and deduced amino acid sequences were analyzed by Omiga and Mega 5.05 software,and a phylogenetic tree was plotted.Results A total of seven CVB5 strains were isolated,of which the homologies of nucleotide and amino acid sequences of partial VP1 genes were 92.7% ～ 98.5% and 93.6% ～ 100%,respectively.The homologies of nucleotide sequences of partial VP1 genes of the seven strains were 80.1% ～ 99.1% to those of other reference strains isolated in China,of which that to strain 98388-SD-CHN-1998 isolated in Shandong Province was low.The homologies of amino acid sequences of partial VP1 genes of the seven strains were 84.4% ～ 100.0% to those of other reference strains isolated in China,of which that to strain CoxB5-Henan-2010 isolated in Henan Province and strain 98388-SD-CHN-1998 isolated in Shandong Province were low.The homologies of nucleotide and amino acid sequences to were 77.5% ～ 89.1% and 92.1% ～ 96.3% to those of reference strains isolated in other countries,and 79.6% ～ 80.9% and 92.1% ～ 95.4% to those of prototype strain Faulkner,respectively.Phylogenetic tree showed that the seven Kunming isolates belonged to a separated branch,while the epidemic strains in China,except 98388-SD-CHN-1998,formed one branch.Conclusion The CVB5 strains isolated in Kunming in 2009 were of the same genetic group.