Objective To analyze the genetic characterization of the VPI gene of Coxsackie B3 virus( named KMA103 - 09) causing meningitis.Methods The specimens of stool were isolated in Hep - 2,RD and Vero cell lines.Viral RNA was extracted and the VP1gene was amplified using RT - PCR and sequenced.The sequence of the VP1 gene was compared with VP1 gene sequences of other Coxsackies virus from Genebank using Mega4.0 software.Results The virUB of Coxsackie B3 virus was confirmed by serum neutralization test.The VP1 gene of Coxsackie virus B3 isolated from stool specimens was 849 nucleotides.There was no missing and insertion of nucleotide.Compared the Coxsackie virus B3 strains with Coxsackie virus B3 strains isolated from provinces in China, the homology of amino acids were 99.2％ - 100％ .The VP1 amino acid sequence was 100% which was identical to that of the Coxsackie virus B3 strain isolated Shandong.The homology was 95.41％ - 96.47％ when compared with the isolatea from other countries.Phylogenetic analysis revealed that the evolutional relationship of the Coxsackie virus B3 was closest to the AM06HZ/SD/CHN/2006/CB3 strains.Conclusion The KMA103 - 09 strain was Coxsackie virus B3.The Vpl gene of Coxsackie virus B3 in china was very conserved.%目的 对一株从无菌性脑膜脑炎患者中分离到的柯萨奇B组3型病毒(coxsackie virus B3,CVB3)分离株KMA103-09进行VP1 区的基因特征分析.方法 采用 Hep-2、RD 细胞对患者粪便标本进行病毒分离,肠道病毒组合血清进行鉴别,反转录 -聚合酶链反应 (RT - PCR)扩增病毒目的 片段,测序后进行VP1基因分析,用 Mega4.1等软件分析处理.结果 肠道病毒组合血清鉴别为柯萨奇病毒B组3 型(Cox.B3),从病毒性脑膜脑炎患者粪便标本中,分离到CVB3,其VP1 区的核苷酸长度为849bp,未发现核苷酸插入与丢失.与山东04433/SD/CHN/2004/CB3株、山东05280/SD/CHN/2005/CB3株及山东YZ127/SD/CHN/2005/CB3株氨基酸同源,与其余国内分离株同源性高于为99.29%,与国外毒株的同源性为95.41%～96.47%.在进化树上与AM06HZ/SD/CHN/2006/CB3株显示在同一个分支上.结论 分离的肠道病毒为柯萨奇病毒B组3型(Cox.B3),分离株(KMA103-09) VP1 区变异较小.
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