摘要目的 对2009年山东省临沂市起无菌性脑膜炎暴发所分离的病原体进行鉴定,并分析其基因特征.方法 采集无菌性脑膜炎患者脑脊液标本42份,分离病毒；RT-PCR扩增阳性分离物,测定VP1核苷酸序列,与GenBank中其他毒株进行同源性分析,并构建VPI基因亲缘进化树.结果 分离到肠道病毒17株,经肠道病毒组合血清中和试验和分子分型鉴定为柯萨奇B5病毒(CVB5).CVB5分离株之间核苷酸和氨基酸同源性分别为92.3％～100.0％和98.7％～100.0％,与CVB5原型株(Faulkner株)的核苷酸和氨基酸的同源性分别为81.0％～82.4％和9 6.6％～97.0％.VP1区亲缘进化树分析显示,全球CVB5可分为A、B、C、D共4个基因型,引起本次无菌性脑膜炎暴发的CVB5属于基因型D,但位于不同的传播链中.结论 CVB5是此次临沂市无菌性脑膜炎暴发的病原体,分离株之间有较大的遗传差异,但均属于D基因型.

abstractsObjective To identify the pathogen of an aseptic meningitis outbreak which occurred in Linyi City of Shandong Province during the summer of 2009,and to analyze the genetic variations of Coxsackicvirus B5 (CVB5) isolates.Methods Forty-two cerebrospinal fluids (CSF) specimens were collected from aseptic meningitis cases and virus isolation was performed. The viral RNA was extracted and amplified from the positive specimens using reverse transcription polymerase chain reaction (RT-PCR).The partial VP1 coding region was purified and sequenced. The phylogenetic trees based on VP1 sequences were constructed among CVB5 isolates and those in GenBank.Results Seventeen enteroviruse strains were isolated from 42 CSF samples with 40.5％ isolation positive rate. All these strains were identified as CVB5 using both microneutralization test and molecular typing methods. Homology comparisons indicated that the nucleotide acid identities and amino acid sequence identities were 92.3 ％- 100.0％ and 98.7 ％- 100.0％,respectively among these CVB5 isolate.s,and compared with the Faulkner prototype strain,which were 81.0％-82.4％ and 96.6％ 97.0％,respectively.Phylogenetic analysis on VP1 sequences showed that all CVB5 could be separated into four genogroups of A,B,C and D.Isolates of this outbreak belonged to genogroup D.Interestingly,two distinct genogroups in the phylogenetic tree were observed among the 17 isolates.Conclusions CVB5 is responsible for the outbreak of aseptic meningitis in Linyi City of Shandong Province,China. The genetic diversity is high among the isolates and all belong to genogroup D.