手足口病柯萨奇病毒A10和A6型的分子鉴定及其VP1区3′端序列分析  

Molecular identification of the pathogens causing HFMD and sequence analysis of VP1 3' terminal region of the Coxsackievirus A10 and A6

作　　者：任倩[1,2] 张乐海[1] 马丽霞[1] 王世富[1] 吕欣[1] 张忠[2] 于爱莲[2] 

机构地区：[1]山东大学齐鲁儿童医院,山东济南250022 [2]泰山医学院病原生物学教研室

出　　处：《中国病原生物学杂志》2011年第8期561-564,570,共5页Journal of Pathogen Biology

摘　　要：目的鉴定手足口病患儿咽拭子病原体柯萨奇病毒A10（CVA10）和A6（CVA6）,并对其VP1编码区3′端序列进行分析。方法合成对肠道病毒VP1区3′端序列具有较高特异性的兼并引物040-011,RT-PCR方法测定其VP1区3′端基因序列,通过BLAST程序比对,鉴定病毒的基因型;与GenBank中其他已知病毒基因型序列进行同源性分析,构建遗传进化树。结果 38例非EV71非CVA16引起的手足口病患儿咽拭子,5例检测到CVA10,1例检测到CVA6。在进化关系上CVA10与D基因型最为密切,核苷酸和氨基酸同源性分别为92.7%~94.2%、91.7%~100%;CVA6与2003~2009年报告的部分毒株核苷酸同源性为86.8%~90.2%,其中与2008年台湾EU908148的毒株同源性最高;氨基酸同源性为85.2%~89.0%。结论兼并引物040-011对CVA10和CVA6有良好的基因扩增特异性;引起手足口病的肠道病毒除EV71和CVA16外,CVA10和CVA6也是重要的病原;鉴于2008年芬兰和2009年山东省文登市CVA10引起的手足口病暴发,应加强对手足口病病原的检测和分型,以了解柯萨奇病毒在手足口病中的位置,掌握其遗传进化特征。Objective To indentify the pathogens Coxsackievirus A10 and A6 from the throat swabs of children with hand,foot,and mouth disease（HFMD） and to analyze the 3＇ terminal sequences of the VP1 coding region.Methods Degenerate primer pair 040-011 is highly specific to the 3＇ terminal sequence of the VP1 region of enterovirus.Primers were synthesized and the 3＇ terminal sequence was detected with RT-PCR.The BLAST procedure was used to identify genotypes and the phylogenetic tree of the virus was constructed based on homology with gene sequences os viruses listed in GenBank.Results Other enteroviruses were detected in 38 children with HFMD not caused by CVA16 or EV71.Results of sequencing and comparison revealed that 5 children had Coxsackievirus A10（CVA10） and one had Coxsackievirus A6（CVA6）.Evolutionarily,CVA10 is closest to genotype D.The homology of nucleotides and amino acids was 92.7%-94.2% and 91.7%-100%,respectively.CVA6 had a nucleotide homology of 86.8%-90.2% with other strains that had been reported in 2003-2009;its highest homology was with EU908148,reported in Taiwan in 2008.CVA6 had an amino acid homology of 85.2%-89.0%.Conclusion The degenerate primer pair 040-011 was highly specific for gene amplification of CVA10 and CVA6.In addition to EV71 and CVA16,CVA10 and CVA6 are also major pathogens causing HFMD.An outbreak of HFMD was reportedly caused by CVA10 in Finland in 2008 and in Wendeng,Shandong Province in 2009.These results indicate that HFMD pathogens must be detected and classified in order to ascertain the epidemiological characteristics and genetic evolution of Coxsackievirus.

关 键 词：手足口病 柯萨奇病毒A10型 柯萨奇病毒A6型 VP1区域 基因序列分析 

分 类 号：R373.23[医药卫生—病原生物学]