北京市CoxA24病毒分子生物学特点的初步研究

A preliminary study on molecular characteristics of CoxA24 isolated in Beijing

目的：了解北京地区引起急性出血性结膜炎的CoxA24病毒的分子生物学特点。方法：收集北京市2007年7月至2007年9月疑似急性出血性结膜炎病例,采集患者眼拭子标本,接种于HeLa细胞,观察细胞是否出现病变。采用逆转录聚合酶链反应法（RT-PCR）对阳性标本进行RNA检测,对RT-PCR结果阳性的标本PCR产物进行克隆测序。结果：检测88例患者眼拭子标本,获得5例阳性样本。将4例阳性的PCR产物进行克隆测序,将获得的4株CoxA24毒株与参考毒株进行核苷酸序列对比分析,结果显示该4株毒株组内核苷酸同源性为98.2%-99.5%,与新加坡提交的毒株同源性为96%-98.2%,与中国杭州和宁波2002年分离的CoxA24毒株其核苷酸同源性为94%-95.7%。系统进化树分析结果表明该4株毒株位于独立的一个分支上。结论：北京地区存在新的CoxA24变异毒株。

Objective：To investigate the molecular characteristics of Coxsackie virus A24 isolated in Beijing.Methods：Conjunctival swab specimens of patients in Beijing during July and September were collected.To isolate virus,each specimen was inoculated for HeLa cells and cells were examined for the appearance of CPE.Positive cells that show CPE were detected by a reverse transcription polymerase chain reaction（RT-PCR）.PCR products were cloned and sequenced.Results：A total of 88 conjunctival swab specimens were examined and 5 positive specimens were detected.Four positive PCR products were sequenced.The four isolated strains shared high homology of 98.2%-99.5%.Compared with the nucleotide sequences of CoxA24 reference strain from GenBank,the highest homology was found between the 4 isolates and the isolates from Singapore,which is 96%-98.2%.Compared with the nucleotide sequences of CoxA24 reference strain from Hangzhou and Ningbo in China in 2002,the homology is 94%-95.7%.Phylogenetic analysis showed that the 4 isolates were located at an independent branch.Conclusion：CoxA24 variants have been found in Beijing,and they are different from the isolates in other cities in China and other countries.