一株引起无菌性脑膜脑炎柯萨奇B组3型病毒的分离和VP1基因分析
投稿时间:2010-12-20  修订日期:2010-12-26  点此下载全文
引用本文:李华,杨卉娟,柯华昕,陈俊英,施海晶,孙强明,马绍辉.一株引起无菌性脑膜脑炎柯萨奇B组3型病毒的分离和VP1基因分析[J].医学研究杂志,2011,40(7):77-80
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作者单位
李华 中国医学科学院/北京协和医学院医学生物学研究所 
杨卉娟 中国医学科学院/北京协和医学院医学生物学研究所 
柯华昕 中国医学科学院/北京协和医学院医学生物学研究所 
陈俊英 中国医学科学院/北京协和医学院医学生物学研究所 
施海晶 中国医学科学院/北京协和医学院医学生物学研究所 
孙强明 中国医学科学院/北京协和医学院医学生物学研究所 
马绍辉 中国医学科学院/北京协和医学院医学生物学研究所 
中文摘要: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。
中文关键词:无菌性脑膜脑炎  柯萨奇病毒B组3型(Cox. B3)  VP1基因
 
Abstract:ObjectiveTo analyze the genetic characterization of the VP1 gene of Coxsackie B3 virus(named KMA103-09) causing meningitis. MethodsThe specimens of stool were isolated in Hep-2,RD and Vero cell lines. Viral RNA was extracted and the VP1 gene 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. ResultsThe virus 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 isolates 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. ConclusionThe KMA103-09 strain was Coxsackie virus B3.The Vp1 gene of Coxsackie virus B3 in china was very conserved.
keywords:Aseptic meningoencephalitis  Coxsackie B3 virus  VP1 gene
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