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研究2010年中国内蒙古自治区引起手足口病(Hand foot and mouth disease,HFMD)的病原谱及人肠道病毒71型(Human enterovirus,HEV71)的分子特征。采集内蒙古自治区12个盟市门诊就诊的HFMD患者粪便和咽拭子标本共921份,进行病毒分离,然后利用三通道实时荧光定量PCR法[同时检测HEV71,柯萨奇病毒A16型(CVA16)和人肠道病毒(HEV)]对阳性分离物进行鉴定,对鉴定为其它HEV的阳性分离物进行VP4和VP1编码区扩增及核苷酸序列测定和分析。921份标本共分离出153株病毒,阳性率为16.61%,其中61株为HEV71,占39.87%,82株为CVA16,占53.59%,7株为其它HEV(分别为6株CVB4和1株Ⅱ型脊髓灰质炎疫苗病毒株),占6.53%,3株为腺病毒。重症病例中分离到9株病毒,其中6株为HEV71,3株为CVA16。选取从临床诊断分别为普通型病例、重型病例的HFMD患者临床标本中分离到的32株HEV71代表株进行VP1编码区基因扩增及核苷酸序列测定和分析,与HEV71其它各基因型和基因亚型的代表株构建亲缘性进化树。32株内蒙古HEV71代表株与1998年以来中国大陆HEV71分离株的VP1区核苷酸和氨基酸水平上的同源性都较高,尤其与2008年的北京代表株同源性最高,与C4基因亚型代表株聚为一支,属于C4基因亚型C4a进化分支,但它们之间的核苷酸和氨基酸的同源性略有差异,分别为96.4%～100%和98.14%～100%,与2007年的内蒙古代表株存在一定的差异,核苷酸同源性为96.95%～97.87%。亲缘进化关系树显示,这些HEV71处于不同的簇中,属于多个病毒传播链。2010年内蒙古HFMD的病原谱以CVA16和HEV71为主,重症病例中以HEV71居多。内蒙古流行的HEV71属于C4基因亚型C4a进化分支,并且存在多个传播链,与2008年北京代表株亲缘关系比2007年内蒙古代表株亲缘关系近,说明内蒙古流行的HEV71不是独立进化的,而是与中国流行的HEV71在共同进化。
To study the molecular pathogens of Hand foot and mouth disease (HFMD) and the molecular characteristics of human enterovirus 71 (HEV71) in Inner Mongolia in 2010. A total of 921 samples of feces and throat swabs from HFMD patients from 12 alliance outpatient clinics in Inner Mongolia Autonomous Region were collected for virus isolation. Three-channel real-time fluorescence quantitative PCR (simultaneous detection of HEV71, Coxsackievirus A16 (CVA16) Human Enterovirus (HEV)] positive isolates were identified, positive isolates identified as other HEVs were amplified for VP4 and VP1 coding region and nucleotide sequence determination and analysis. A total of 153 strains of viruses were isolated from 921 samples, with a positive rate of 16.61%, of which 61 were HEV71 (39.87%), 82 were CVA16 (53.59%) and 7 were other HEVs (6 CVB4 and 1 strain respectively) Polio vaccine strain), accounting for 6.53%, three strains of adenovirus. Nine strains of viruses were isolated from severe cases, of which 6 were HEV71 and 3 were CVA16. 32 strains of HEV71 isolated from clinical samples of HFMD patients with clinically diagnosed common and severe cases respectively were used to amplify and analyze the nucleotide sequence of the VP1 coding region and to compare with other HEV71 genotypes and genes Subtype representative strains construct phylogenetic tree. The 32 representative strains of HEV71 in Inner Mongolia had higher nucleotide and amino acid homology with the VP1 region of HEV71 isolates in mainland China since 1998, especially those homologous with Beijing representative strains in 2008 and C4 genes Type represented a cluster of C4 subtypes belonging to the C4a evolutionary branch, but their nucleotide and amino acid homologies were slightly different, ranging from 96.4% to 100% and 98.14% to 100%, respectively There was a certain difference between representative strains of Inner Mongolia in 2007, with nucleotide homology of 96.95% -97.87%. Phylogenetic tree shows that these HEV71s are in different clusters and belong to more than one virus transmission chain. In 2010, the epidemic spectrum of HFMD in Inner Mongolia was dominated by CVA16 and HEV71, and HEV71 was predominant in severe cases. Inner Mongolia popular HEV71 belongs to the C4 gene subtype C4a evolutionary branch, and there are multiple transmission chain, and Beijing representative strains in 2008 relatives than in 2007, the Inner Mongolia representative strains of kinship, indicating that epidemic HEV71 in Inner Mongolia is not independent evolution, but HEV71 popular with China is co-evolving.