济南市轻重症手足口病患儿肠道病毒71型结构蛋白基因特征分析  

Sequence analysis of structure gene of enterovirus 71 strains isolated from mild and severe HFMD cases in Jinan City

作　　者：赵怀龙[1] 王春荣[1] 杨国樑[1] 赵宝添 吕燕[1] 刘岚铮[1] ZHAO Huai-long;WANG Chun-rong;YANG Guo-liang;ZHAO Bao-tian;LV Yan;LIU Lan-zheng(Jinan Municipal Center for Disease Control and Prevention,Jinan 250021,China)

机构地区：[1]济南市疾病预防控制中心,山东济南250021

出　　处：《现代预防医学》2020年第10期1861-1864,1887,共5页Modern Preventive Medicine

基　　金：山东省医药卫生科技发展计划项目(2017WS529)。

摘　　要：目的比较分析济南市轻重症手足口病(hand,foot and mouth disease,HFMD)病原肠道病毒71型(enterovirus 71,EV71)结构蛋白基因及氨基酸变异情况。方法对2017年济南市轻重症手足口病患儿EV71细胞分离株结构蛋白基因序列进行RT-PCR扩增测序,应用EditSeq和MegAlign等软件对结构蛋白基因进行序列拼接及核苷酸、氨基酸同源性比对,应用Mega 5.2软件构建VP1基因系统进化树。结果轻重症患儿EV71分离株结构蛋白基因核苷酸和氨基酸同源性分别为93.0%~98.8%和99.0%~99.8%,均属于C4a基因亚型。经VP1基因同源性分析,济南市EV71分离株与2017年江苏株(GenBank ID:MG520666,同源性99.3%)、2015年河南株(KU744452,同源性99.2%)、2015年北京株(KU376386,同源性99.2%)及2016年山东潍坊株(MG588036,同源性99.0%)亲缘关系较近。死亡患儿EV71分离株VP1蛋白出现E145Q突变,重症EV71分离株分别出现VP1蛋白A19V、T101I,VP3蛋白M150I突变。结论济南市轻重症HFMD患儿EV71分离株均属于C4a亚型,与江苏、河南、山东及北京等地的EV71株亲缘关系较近,其VP1蛋白145位氨基酸残基可能是影响病毒毒力的重要位点。Objective To investigate the structural protein gene characteristics and amino acid variation of enterovirus 71(EV71)isolated from the mild and severe Hand-foot-and-mouth Disease(HFMD)cases in Jinan.Methods The EV71 strains isolated from mild and severe HFMD cases in Jinan city in 2017 were amplified the sequences of structural protein gene using reverse transcription-polymerase chain reaction(RT-PCR)method.All the structural protein genes were spliced and analyzed for the homology consistency of nucleotide and amino acid with the EditSeq and MegAlign software,and VP1gene phylogenetic tree was constructed with the MEGA5.2 software.Results The homogeneity between the EV71 strains isolated from mild and severe HFMD patients were 93.0%-98.8%and 99.0%-99.8%respectively for the structural protein gene and deduced amino acid.All the EV71 strains isolated from Jinan were belonged to C4a sub-genotype,According to the homogeneity analyses of VP1 gene,Jinan EV71 strains were closely related to strains detected in Jiangsu(GenBank accession number MG520666),Henan(GenBank accession number KU744452),Beijing(GenBank accession number KU376386)and Shandong(GenBank accession number MG588036).Three Jinan EV71 strains originated from severe cases had amino acid substitution,such as E145Q,A19V,T101I in VP1 protein,and M150I in VP3 protein.Conclusion The EV71 strains isolated from Jinan belong to C4a sub-genotypes and share high homology with the strains isolated form Jiangsu,Henan,Shandong and Beijing areas.VP1-145 site of EV71 is a potential neurovirulence determinant.

关 键 词：手足口病 肠道病毒71型 结构基因 系统进化树 

分 类 号：R117[医药卫生—公共卫生与预防医学]