临沂市脑膜炎／脑炎症候群监测病例埃可病毒30型的分离及其基因特征分析  

Genetic analysis of echovirus 30 isolated from acute meningitis/encephalitis syndrome cases in Linyi city of Shandong province, China

作　　者：杨静[1] 张晓丽[2] 陶泽新[3] 王海岩[3] 林小娟[3] 季圣翔[2] 刘尧[1] 张玉军[2] 徐爱强[3] 

机构地区：[1]山东大学公共卫生学院流行病与卫生统计学研究所,济南250012 [2]临沂市疾病预防控制中心 [3]山东省疾病预防控制中心山东省传染病预防控制重点实验室

出　　处：《中华实验和临床病毒学杂志》2011年第6期409-412,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：山东省医药卫生科技发展计划项目（2011HZ058、2011QZ013）

摘　　要：目的 调查山东省临沂市脑膜炎/脑炎症候群（AMES）病例中人类肠道病毒病原谱的构成,分析主要病原体埃可病毒30型的基因特征.方法 采集AMES病例的脑脊液、粪便和（或）咽拭子标本,所有标本经预处理后,接种RD、Hep-2细胞分离病毒,阳性分离物采用血清学和分子生物学方法鉴定型别,与GenBank检索到的其他Echo30毒株进行同源性比较,并构建VP1完整编码区系统进化树研究其遗传进化规律.结果 2010年4-12月,共采集101例病例的222份标本（脑脊液56份,咽拭子94份,粪便72份）,其中34例患者的47份标本（脑脊液4份,咽拭子18份,粪便25份）中分离到HEV,11例鉴定为Echo30.通过同源性分析,发现其核苷酸同源性与2003年山东省泰安市、章丘市及江苏省分离株最近（91.2％～95.4％）,与原型株差异较大（17.9％～19.6％）；11株临沂分离株之间核苷酸差异为0～10.2％.系统进化树分析显示,临沂分离株属于基因型6的3个不同分支,提示来自3个传播链的Echo30病毒在临沂市共同循环.结论 山东省临沂市2010年AMES病例标本的优势分离株为Echo30,与国外分离株属于不同的进化分支,具有不同的基因特征.Objective To investigate the pathogen spectrum constitution of HEV related to acute meningitis/encephalitis syndrome（AMES） and to analyze the genetic characterization of the echovirus 30 （Echo30） isolates.Methods Cerebrospinal fluid,stool,and/or throat swab specimens from 101 AMES cases were collected for virus isolation with RD and HEp-2 cell lines in Linyi city from April to December,2010.After typing by neutralization test,the entire VP1 gene of the isolates were amplified by RT-PCR and sequenced.Homologous comparison was carried out and phylogenetic analysis were performed.Results HEVs were isolated from 47 specimens of 34 patients（4 from cerebrospinal fluid,18 from throut swab and 25 from stool）,with 11 Echo30 isolates identified by microneutralization assay and molecular typing method.In alignment of VP1 sequences with global Echo30 isolates,relative high nucleotide homologies（91.2% -95.4% ） with Taian,Zhangqiu and Jiangsu isolates and high divergences（ 17.9% - 19.6% ） with prototype strain were observed.Nucleotide divergences among Linyi isolates were 0 - 10.2%,and 3 lineages were revealed via phylogenetic analysis,reflecting 3 transmission chains of Echo30 co-circulated in Linyi city,2010.Conclusion Echo30 was the predominant serotype of AMES cases in Linyi city,2010.These isolates possessed considerable divergence with Echo30 from other countries.

关 键 词：脑膜炎-病毒性 脑炎 病毒性 肠道病毒B型 人 种系发生 

分 类 号：R51[医药卫生—内科学]