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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 evolutionary characteristics of Shandong local strains of enterovirus A group, we collected fecal samples from patients with acute flaccid paralysis (AFP) and hand, foot and mouth disease (HFMD) in 2008 and 2009 from 1998 to 2008, Enterovirus separation. Positive virus isolates from HFMD cases were identified by Enterovirus 71 (EV71) and Coxsackie virus A16 (CVA16) -specific reverse transcription-polymerase chain reaction, and then some positive isolates were selected for VP1 complete coding region Nucleotide sequence determination and analysis. The EV71 and CVA16-specific primer-negative and positive virus isolates from AFP cases were tested for the nucleotide sequence of VP1 complete coding region using group A enterovirus-specific primers. The obtained sequences were compared with those of other A group representative genotypes and the phylogenetic tree was constructed. A total of 293 strains of enterovirus A Shandong were obtained, belonging to 8 genotypes, CVA4, CVA5, CVA10, CVA14, CVA16, EV71, EV76 and EV90 respectively. Shandong province isolates in each genotype had lower homology with the original strains. Phylogenetic analysis showed that the serotypes of Shandong local strains of enterovirus A were distantly related to the prototype strains, suggesting the existence of Shandong province Group A intestine virus transmission with different genetic characteristics.