河南省濮阳市7株肠道病毒71型全基因组序列分析  

Complete genome sequencing of 7 strains of enterovirus 71 from the City of Puyang,Henan Province

作　　者：徐国锋[1] 王瑞[1] 张清杰[1] 张玉华[1] 孟晓飞[1] 隋美丽[2] 李懿[3] 卫海燕[3] 黄学勇[3] 

机构地区：[1]濮阳市疾病预防控制中心,河南濮阳457000 [2]郑州大学公共卫生学院,河南郑州450001 [3]河南省疾病预防控制中心,河南郑州450016

出　　处：《中国病原生物学杂志》2016年第7期607-610,619,共5页Journal of Pathogen Biology

基　　金：河南省科技攻关计划项目(No.122102310268);河南省医学科技重大攻关项目(No.201001015);河南省卫生科技创新型人才工程中青年科技创新人才项目(No.cxyx20130406)

摘　　要：目的了解濮阳地区手足口病患者标本中分离的肠道病毒71型(EV71)病毒全基因组特征。方法选取本市发生的7例手足口病患者肠道病毒EV71型核酸阳性标本,使用人横纹肌肉瘤细胞(rhabdomyosarcom cell,RD)进行病毒培养,RT-PCR型别鉴定,对分离的7株EV71型毒株通过12对引物进行分段扩增,扩增产物进行EV71型全基因组序列测定和基因进化特性分析,利用生物信息学软件对基因组序列进行分析并构建EV71病毒分类和发育图谱。结果共分离出7株EV71病毒,系统发育树显示7株病毒在分类上均属于C4亚型C4a分支。在发育相似度上主要分为两大类,其中PY464与宁波2010年NINGBO.CHN/065/2010EV71分离株相似度最高;其他6株呈现聚类,与山东临沂EV71SDLY107分离株相似度最高。在同一时期、同一地区,轻、重症病例分离株VP1区段无明显差别,而全基因区段存在一定差别。结论在濮阳市同一地区、同一时期,亦呈现出以某一种手足口病病毒为主的流行趋势。Objective To ascertain the complete genome sequence of enterovirus 71（EV71）isolated from patients with hand,foot,and mouth disease（HFMD）in the City of Puyang. Methods Seven strains of enterovirus 71 were isolated in the City of Puyang.The virus was cultured in RD cells and identified using RT-PCR.Twelve pairs of primers were used to amplify the 7EV71 strains,and the whole genome sequence and genetic evolutionary characteristics of EV71 were also analyzed.Bioinformatics was used to analyze the genome sequence and to classify and chart the development of the EV71 virus in the Puyang region. Results Seven EV71 strains were isolated.A phylogenetic tree was constructed for the 7EV71 strains and it indicated that all 7EV71 strains belonged to subtype C4 a.All 7strains were divided into two categories.The PY464 strain displayed the greatest similarity to the NINGBO.CHN/065/2010 isolate of EV71 while the other 6strains formed a cluster that displayed the greatest similarity to the SDLY107 isolate from the City of Linyi,Shandong Province.There were no differences in the VP1 region of strains isolated from patients with a severe or mild case HFMD in the same area and at the same time,but certain differences in complete genome sequences of those strains did exist. Conclusion The predominant pathogen that caused HFMD displayed certain trends in that a certain virus was predominant in the same area and at the same time in the City of Puyang.

关 键 词：肠道病毒71型 全基因测序 系统发育 

分 类 号：R373.2[医药卫生—病原生物学]