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在前期研究中,对山东省1989～2011年急性弛缓性麻痹(Acute flaccid paralysis,AFP)和手足口病(Hand,foot and mouth disease,HFMD)患者标本中分离到的部分人类肠道病毒(Human enteroviruses,HEVs)进行分子定型,发现了8个HEV-A血清型。为进一步探究HEV-A组病毒在山东省的基因型分布及分子进化特征,本研究继续对剩余分离株进行核酸提取,以HEV-A组特异性引物对VP1完整编码区进行序列扩增、测序和分子定型,又鉴定出4个血清型,分别为柯萨奇病毒A组(Coxsackievirus A,CVA)2、6、8、12型,共7株病毒。同源性分析显示,7株HEV-A山东分离株与其原型株核苷酸同源性介于80.8%～85.0%之间。系统进化树表明,CVA8、12型山东分离株与国内分离株存在一定亲缘关系,而CVA2、6型山东株同国内外分离株亲缘关系均较远,并且CVA2、6型在山东省内可能存在多个传播链。本研究将我省HEV-A组的血清型别增加至12个,进一步丰富了HEV-A山东地方株的遗传进化特征,并且发现CVA2、6、8、12血清型在山东省甚至全国范围内存在不同基因特征的传播链的共循环。
In the previous study, some human enteroviruses isolated from specimens of patients with acute flaccid paralysis (AFP) and hand, foot and mouth disease (HFMD) from 1989 to 2011 in Shandong Province enteroviruses, HEVs) and found 8 HEV-A serotypes. In order to further explore the genotype distribution and molecular evolution characteristics of HEV-A virus in Shandong province, we continued to extract nucleic acids from the remaining isolates. The complete coding region of VP1 was amplified by HEV-A group specific primers and sequenced And molecular typing. Four serotypes were identified, which were Coxsackievirus A (CVA) type 2, 6, 8, and 12 viruses, respectively. Homology analysis showed that the nucleotide homology of seven HEV-A Shandong isolates and their prototype strains ranged from 80.8% to 85.0%. The phylogenetic tree showed that there was a certain genetic relationship between CVA8 and Shandong isolates 12 and domestic isolates, whereas CVA2 and Shandong isolates 6 were far away from domestic and foreign isolates, and CVA2 and 6 were likely to exist in Shandong province Multiple communication chains. This study increased the serotypes of HEV-A group in our province to 12, further enriching the genetic evolutionary characteristics of HEV-A Shandong local strains and found that CVA2, 6, 8 and 12 serotypes were still existed in Shandong province and even nationwide Circulation of different strands in different gene traits.