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[2010年内蒙古自治区手足口病致病谱及肠道病毒71型分子特征]

[The pathogenic spectrum of hand, foot and mouth disease and molecular characterizations of human enterovirus 71 in Inner Mongolia autonomous region in 2010].

作者信息

Tian Xiao-Ling, Zhang Yong, Yan Shao-Hong, Ma Xue-En, Wang Wen-Rui

机构信息

College of Veterinary Medicine, Inner Mongolia Agricultural University, Huhhot 010018, China.

出版信息

Bing Du Xue Bao. 2013 May;29(3):304-9.

Abstract

To study the pathogenic spectrum of hand, foot and mouth disease (HFMD) and the molecular characterizations of human enteroviruses 71 (HEV71) isolated from the clinical specimens of HFMD patients in Inner Mongolia in 2010. A total of 921 clinical specimens including stools and throat swabs were collected from HFMD patients in outpatient service in Inner Mongolia and then viral isolation was performed, the positive viral isolates were identified by using the real-time PCR method (detecting EV, HEV71 and CVA16 in a single tube), and VP4 and VP1 coding region amplification and sequencing was performed with the viral isolates that were identified as non-HEV71, non-CVA16 HEVs. A total of 153 viruses were isolated form 921 clinical specimens, the positive rate was 16.61%, of which 61 (39.87%) were HEV71, 82 (53.59%) were CVA16, 7 (6.53%) were other HEVs(6 were CVB4 and 1 was polio vaccine virus type II) and 3 (1.96%) were adenoviruses. Nine viruses were isolated from severe cases, of which 6 were HEV71 and 3 were CVA16. Thirty two HEV71 isolates were selected from the patients presenting mild symptoms and the patients presenting severe symptoms randomly, and the VP1 coding regions of represented HEV71 isolates were amplified and sequenced. Finally the phylogenetic tree was constructed among the VP1 coding regions of the different genotypes and subgenotypes of HEV71 strains. The nucleotide acid and amino acid of 32 represented HEV71 strains in Inner Mongolia were closed to HEV71 strains isolated from mainland China since 2007, especially from Beijing in 2008, and it showed that all HEV71 strains clustered within the C4a evolution branch of C4 subgenotype. There was slight difference in the nucleotide and the amino acid sequence in VP1 region among the 32 Inner Mongolia HEV71 strains, the identity were 96.4%-100% and 98.14%-100%, respectively, and there was a little difference in the nucleotide acid sequence between the HEV71 strains from Inner Mongolia in 2010 and in 2007, the identity was from 96.95% to 97.87%. Thirty two HEV71 strains were in different lineages in the phylogenetic tree, and it indicated that these strains belonged to many different viral transmission chains. HEV71 and CVA16 were the main pathogens of HFMD in Inner Mongolia in 2010 and most severe cases were caused by HEV71. All the HEV71 strains circulated in Inner Mongolia belonged to C4a evolution branch within C4 subgenotype. Phylogenetic analysis revealed that 2010 Inner Mongolia HEV71 strains were located in different lineages, and had more nucleotide identity with 2008 Beijing HEV71 strains than with 2007 Inner Mongolia HEV71 strains. This indicated that Inner Mongolia HEV71 strains had not evolved independently, but co-evolved with the HEV71 strains in other provinces in mainland China.

摘要

研究2010年内蒙古手足口病(HFMD)的病原谱及从手足口病患者临床标本中分离出的肠道病毒71型(HEV71)的分子特征。从内蒙古门诊的手足口病患者中收集了包括粪便和咽拭子在内的921份临床标本,随后进行病毒分离,采用实时荧光定量PCR方法(单管检测肠道病毒、HEV71和柯萨奇病毒A16型(CVA16))对阳性病毒分离株进行鉴定,对鉴定为非HEV71、非CVA16的肠道病毒的病毒分离株进行VP4和VP1编码区扩增及测序。从921份临床标本中共分离出153株病毒,阳性率为16.61%,其中61株(39.87%)为HEV71,82株(53.59%)为CVA16,7株(6.53%)为其他肠道病毒(6株为柯萨奇病毒B4型(CVB4),1株为Ⅱ型脊髓灰质炎疫苗病毒),3株(1.96%)为腺病毒。从重症病例中分离出9株病毒,其中6株为HEV71,3株为CVA16。从表现为轻症和重症的患者中随机选取32株HEV71分离株,对代表性HEV71分离株的VP1编码区进行扩增及测序。最后构建了HEV71毒株不同基因型和亚基因型VP1编码区的系统进化树。内蒙古32株代表性HEV71毒株的核苷酸和氨基酸与2007年以来从中国大陆分离的HEV71毒株相近,尤其是与2008年北京的毒株相近,表明所有HEV71毒株均聚集在C4亚基因型的C4a进化分支内。32株内蒙古HEV71毒株VP1区核苷酸和氨基酸序列存在细微差异,同一性分别为96.4% - 100%和98.14% - 100%,2010年与2007年内蒙古HEV71毒株的核苷酸序列也存在少许差异,同一性为96.95%至97.87%。32株HEV71毒株在系统进化树中处于不同分支,表明这些毒株属于多个不同的病毒传播链。HEV71和CVA16是2010年内蒙古手足口病的主要病原体,大多数重症病例由HEV71引起。在内蒙古流行的所有HEV71毒株均属于C4亚基因型内的C4a进化分支。系统进化分析显示,2010年内蒙古HEV71毒株处于不同分支,与2008年北京HEV71毒株的核苷酸同一性高于与2007年内蒙古HEV71毒株的核苷酸同一性。这表明内蒙古HEV71毒株并非独立进化,而是与中国大陆其他省份的HEV71毒株共同进化。

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