一株柯萨奇病毒B组5型( Cox. B5)病毒的分离及VP1基因分析

摘要

Objective To analyze the genetic characterization of the VP1 gene of coxsackie virus B5 causing meningitis isolated from Kunming in 2009. Methods The specimens of stool were isolated in Hep -2 and RD 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 coxsackie virus from Genbank using Mega4.0 software. Results The VP1 gene of coxsackie virus B5 isolated from stool specimens was 831 nucleotides. There was no missing and insertion of nucleotide. Compared with the coxsackie virus B5 strains isolated from other countries, the homology of amino acids were 95. 67% -98.19% . Phylogenetic analysis revealed that the evolutional relationship of the coxsackie virus B5 was closest to the YZ081/SD/CHN/2005/CB5 strains. Conclusion The KMA193 -09 strain was belong to coxsackie virus B5. The VP1 gene of coxsackie virus B5 in china was strictly conserved.%目的 研究2009年昆明市无菌性脑膜炎的病原柯萨奇B5( coxsackie virus B5,CVB5)分离株(KMA193 - 09)的VP1基因特征.方法 采用RD细胞、Hep -2细胞对患者粪便标本进行病毒分离,反转录-聚合酶链反应(RT - PCR)扩增病毒VP1基因并进行序列测定,用Mega 4.0等软件分析处理.结果 从无菌性脑膜炎患者粪便标本中,分离到CVB5,其VP1区的核苷酸长度均为831bp,未发现核苷酸插入与丢失.与浙江COXB5/ZHEJIANG/12/02 (CFS)株、山东02336/SD/CHN/2002/C B5株及浙江COXB5/ZHEJIANG/13/02株氨基酸同源性最高为98.19％,与国外毒株的同源性为95.67％～97.83％.在进化树上与YZ081/SD/CHN/2005/CB5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5型(CVB5),分离株(KMA193-09) VP1区变异较小.