2009年深圳地区肠道病毒71型毒株VP4基因进化分析  

2009 evolution for VP4 of enterovirus 71 strains in Shenzhen

作　　者：冼慧霞[1] 陈龙[1] 阳帆[1] 罗敏[1] 吴春利[1] 杨洪[1] 张海龙[1] 姚相杰[1] 何雅青[1] 

机构地区：[1]深圳市疾病预防控制中心重大传染病监控实验室

出　　处：《中华实验和临床病毒学杂志》2013年第1期11-13,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：国家自然科学基金资助（81273075,2012）

摘　　要：目的对深圳地区手足口病（hand，foot and mouth disease，HFMD）肠道病毒71型分离株（enterovirus71，EV71）的VP4基因遗传进化进行分析。方法2009年采集深圳市儿童医院就诊的HFMD患者的粪便标本491份，选取其中8份经细胞培养鉴定为阳性的EV71毒株用RT-PCR扩增VP4基因，利用MEGA4．0软件进行遗传进化分析。结果2009年深圳地区8株EV71VP4基因全长207bp，编码69个氨基酸；8株EV71VP4基因核苷酸同源性为94．2％-98．1％，与深圳2001至2004年分离的17株EV71毒株核苷酸同源性在89．9％～98．1％，与GenBank中其他EV71病毒株VP4的核苷酸同源性为79．2％-100％；其中与亚洲流行株阜阳株（EU703812）核苷酸的同源性最高（100％），其次是C4代表株及2004年深圳株（AY895144）为94．2％～97．1％。除了印度株和其中1株EV71的VP4编码的氨基酸在第54位（ACA）不同之外，其余7株EV71VP4编码的氨基酸序列之间以及与GenBank报道的其他序列同源性达100％。8株EV71VP4基因核苷酸与c4代表株相比有17处不同，除1处以外其余全在简并密码位点上；轻重症病例毒株之间VP4基因序列未见明显改变。进化树显示8株EV71均属于C4基因亚型。结论2009年深圳市流行的EV71属于C4基因亚型，流行的EV71VP4基因非常保守，不属于变异区段，绝大部分核苷酸的变异属无义突变，VP4基因编码的氨基酸变异几乎为0。Objective To analyze the genetic evolution for the common causative agent of hand,foot and mouth disease（HFMD） that VP4 of human enterovirus 71 in Shenzhen district. Method 491 stool specimen were collected from children with hand, foot and mouth diease in Shenzhen Children＇s Hospital 2009. After cell culture, VP4 gene of eight EV71 strains were amplified by reverse-transcriptase PCR（ RT- PCR）, phylogenetic analysis of the VP4 gene was constructed by using MEGA 4.0. Result The VP4 gene of eight EV71 strains encoding 69 amino acids with full length 207 bp. The nucleotide homology of VP4 gene among eight EV71 strains was 94.2%-98.1% , compared with VP4 gene of EV71 strains retrieved from Shenzhen 2001 to 2004 and GenBank was 89.1%-98.1% and 79.2%-100% respectively. Asian epidemic strain Fuyang had the highest nucleotide homology, representative strain C4 and Shenzhen strain （AY895144） with 94. 2% -98. 1% secondly. Except for the 54th amino acid of VP4 gene of India reported strain and one of the eight EV71 strains, the homology of the rest amino acids between the eight EV71 strains and those in GenBank was 100%. Compared with representative strain C4,there were seventeen differences in nueleotide sequences of VP4 of the eight EV71 strains. All of the different nucleotides were located at the degenerate password sites except one. There was no significant difference in VP4 gene between the severe and the mild cases of strains. The eight Shenzhen EV71 strains were classified as sub-genotype C4 in the phylogenetic tree. Conclusions The epidemic of EV71 in Shenzhen 2009 was sub-genotype C4. VP4 gene of EV71 was very conservative which dose not belong to the variation section. The variation of most of nueleotide was invalid variation. The amino acids encoded by VP4 gene which variation was almost zero.

关 键 词：肠道病毒属 基因 种系发生 

分 类 号：R72[医药卫生—儿科]