[2008年北京市手足口病患者肠道病毒71型VP1序列的遗传分析]。

中国疫苗和免疫 Pub Date : 2009-12-01
Fang Huang, Wei-hong Li, Xiao-juan Tan
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引用次数: 0

摘要

目的:获取2008年北京地区HEV71病毒VP1编码区的遗传信息。方法:用横纹肌肉瘤(rhabdomyosarcoma, RD)细胞在33例手足口病患者发病后3 d内采集咽拭子标本,分离肠病毒,采用RT-PCR方法,用特异性引物对人肠病毒进行鉴定,扩增VP1编码区,用Sanger双脱氧法对VP1编码区进行测序。采用Bioedit 7.0.5和MEGA3.1进行核苷酸序列比对和系统发育分析。结果:从33份样本中分离到16株病毒,其中鉴定为HEV71型14株,CVA16型2株,CVA16和HEV71型共感染1株。对10株HEV71分离株VP1核苷酸序列进行序列分析,同源性分析结果表明,VP1核苷酸同源性分别为95.5% ~ 100%和98.9% ~ 100%;2008年阜阳代表性菌株,核苷酸识别率为95.4% ~ 99.1%;代表性菌株C4核苷酸同源性在92%以上。对HEV71株VP1编码区核苷酸序列的系统发育分析表明,北京地区HEV71株属C4亚基因型C4a聚类,10株形成4个相对独立的聚类。结论:北京市手足口病患儿分离的HEV71病毒属C4亚基因型,2004年以来以C4a聚集型为主。系统发育分析表明,2008年北京市HEV71病毒共分属于4个不同聚集群。由于HEV71病毒在中国连续大规模流行,急需收集更多的病毒学建议和HEV71分子流行病学信息。
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[Genetic analysis of VP1 sequences of enterovirus 71 isolated from patients of hand, foot and mouth disease in Beijing, 2008].

Objective: To get genetic information of VP1 coding region of HEV71 in Beijing in 2008.

Methods: 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.

Results: 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.

Conclusions: 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.

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