Huang Fang, Li Wei-hong, Tan Xiao-juan
Beijing Municipal Center for Diseases Control and Prevention, Beijing 100013, China.
Zhongguo Yi Miao He Mian Yi. 2009 Dec;15(6):527-30.
To get genetic information of VP1 coding region of HEV71 in Beijing in 2008.
Enteroviruses were isolated from samples of throat swabs collected from 33 HFMD patients within 3 days after onset by rhabdomyosarcoma(RD) cells and identified by RT-PCR method with specific primers to human enteroviruses, then VP1 coding region was amplified and sequenced by Sanger dideoxy. Bioedit 7.0.5 and MEGA3.1 were used for the nucleotide sequence alignment and phylogenetic analysis.
16 virus strains were isolated from 33 samples, of which 14 strains were identified as HEV71 and 2 were CVA16, and 1 was co-infected with CVA16 and HEV71. Sequence analysis of VP1 nucleotide sequences of 10 HEV71 isolates showed that homologous analysis of nucleotide identity amino acid 95.5%-100% and 98.9%-100% respectively; representative strains of Fuyang in 2008, nucleotide identity was 95.4%-99.1%; with representative strain of C4 nucleotide identity was over 92%. Phylogenetic analysis of HEV71 strains for the nucleotide sequence of VP1 coding region clarified that the HEV71 isolates in Beijing belonged to C4a cluster of C4 sub-genotype and 10 strains formed four relatively separated clusters.
The HEV71 viruses isolated from children of HFMD in Beijing belonged to C4 sub-genotype, and C4a cluster which was the predominant in China since 2004. According to phylogenetic analysis, HEV71 which belonged to more than 4 different clusters were circulating in Beijing in 2008. More virological suggestion for disease control and prevention, and information of HEV71 molecular epidemiology need to be collected urgently due to the successive large epidemic of HEV71 in China.
获取2008年北京地区肠道病毒71型（HEV71）VP1编码区的基因信息。
采用横纹肌肉瘤（RD）细胞，从33例手足口病（HFMD）患者发病3天内采集的咽拭子样本中分离肠道病毒，并用针对人肠道病毒的特异性引物通过RT-PCR方法进行鉴定，然后用桑格双脱氧法扩增VP1编码区并测序。使用Bioedit 7.0.5和MEGA3.1进行核苷酸序列比对和系统发育分析。
从33份样本中分离出16株病毒，其中14株鉴定为HEV71，2株为柯萨奇病毒A16型（CVA16），1株为CVA16和HEV71混合感染。对10株HEV71分离株的VP1核苷酸序列分析显示，核苷酸同源性分析氨基酸分别为95.5%-100%和98.9%-100%；2008年阜阳的代表性毒株，核苷酸同源性为95.4%-99.1%；与C4代表性毒株核苷酸同源性超过92%。对HEV71毒株VP1编码区核苷酸序列的系统发育分析表明，北京地区的HEV71分离株属于C4亚型的C4a簇，10株形成4个相对分离的簇。
从北京地区手足口病患儿中分离出的HEV71病毒属于C4亚型，且为2004年以来在中国占主导地位的C4a簇。根据系统发育分析，2008年北京地区有4种以上不同簇的HEV71在流行。鉴于中国HEV71连续大规模流行，迫切需要收集更多病毒学信息以用于疾病防控及HEV71分子流行病学研究。
