肠道病毒A组山东地方株的基因型分布

Genotype Distribution of Enterovirus A Species Isolated in Shandong Province,China

为研究肠道病毒A组山东地方株的基因型及分子进化特征,本研究采集1998-2008年急性弛缓性麻痹（AFP）患者及2008年和2009年手足口病（HFMD）患者粪便标本,进行肠道病毒分离。来自HFMD病例的阳性病毒分离物进行肠道病毒71型（EV71）和柯萨奇病毒A16型（CVA16）特异性逆转录-聚合酶链反应鉴定,随即选取部分阳性分离株进行VP1完整编码区核苷酸序列测定和分析。EV71和CVA16特异性引物阴性者以及来自AFP病例的阳性病毒分离物采用A组肠道病毒特异性引物进行VP1完整编码区核苷酸序列测定。所获得序列与其他A组各基因型代表株进行比较并构建系统发生树。共获得293株肠道病毒A组山东地方株,分别属于CVA4、CVA5、CVA10、CVA14、CVA16、EV71、EV76、EV90共8个基因型。各基因型内山东地方分离株跟原型株相比基因序列同源性较低,系统发生学分析显示肠道病毒A组山东地方株各血清型中与原型株亲缘关系较远,提示山东省存在不同基因特征的A组肠道病毒传播。

In order to study the genotypes and molecular evolution of human enterovirus （HEV） A species in Shandong Province, Stool samples were collected from AFP and HFMD patients in Shandong Province and virus isolation was performed. Reverse Transeription-Polymerase Chain Reactions （RT-PCR） specific for EV71 and CVA16 were performed with the virus isolates from HFMD patients. Positive isolates were selected for entire VP1 coding gene amplification and sequencing. Isolates with negative PCR results and isolates from AFP patients were selected for entire VP1 coding gene amplification and sequencing using primers specific for HEV A species. Phylogenetic tree was constructed among these VP1 nucleotide sequences and of other strains. Altogether 293 strains classified into 8 genotypes were isolated. The homologous comparison and phylogenetic analysis showed Shandong strains were distinct with prototype strains in every genotype. This report presents an overview of HEV-A in Shandong Province.