2008年和2010年肠道病毒7l型广州分离株全基因组序列分析

钟家禹; 朱冰; 华亮; 王长兵; 邝璐; 谢嘉慧; 陈翊

Abstract

钟家禹,朱冰,华亮,王长兵,邝璐,谢嘉慧,陈翊.2008年和2010年肠道病毒7l型广州分离株全基因组序列分析[J].Chinese journal of Epidemiology,2011,32(7):700-704

2008年和2010年肠道病毒7l型广州分离株全基因组序列分析

Complete genomic sequence analysis on human enterovirus 71 strain in Guangzhou，in 2008 and 20lO

Received:January 26, 2011

DOI：

KeyWord: 道病毒71 基因组序列分析点突变

English Key Word: Enterovirus 7l Genome Sequence analysis Point mutation

FundProject:国家自然科学基金(30972630);广东省自然科学基金(10151012001000002)

Author Name	Affiliation	E-mail
ZHONG Jia-yu	Gnangzhou Women and Children’S Medical Center, Guangzhou 510120, China
ZHU Bing	Gnangzhou Women and Children’S Medical Center, Guangzhou 510120, China	zhubin90327@yahoo．com．cn
HUA Liang	Gnangzhou Women and Children’S Medical Center, Guangzhou 510120, China
WANG Chang-bing	Gnangzhou Women and Children’S Medical Center, Guangzhou 510120, China
KUANG Lu	Gnangzhou Women and Children’S Medical Center, Guangzhou 510120, China
XIE Jia-hui	Gnangzhou Women and Children’S Medical Center, Guangzhou 510120, China
CHEN Yi	Gnangzhou Women and Children’S Medical Center, Guangzhou 510120, China

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Abstract:

目的研究2008年和2010年广州地区手足口病患者中肠道病毒71型(EV71)病毒全基冈组序列的基因型与变异特点。方法参照GenBank上EV71深圳株SHZH03(AY465356)基因组设计分段扩增引物，进行RT-PCR分段扩增EV71病毒基因组，PCR产物直接进行序列测定，用ClustalW／X、DNASTAR、MEGA4．1等软件分析基因组序列。结果克隆9株广州株EV71，病毒全基因组全长序列均为7405 bp，提交到GenBank上的序列号为HQ456305、HQ456306、HQ456307、HQ456308、HQ456309、HQ456310、HQ4563ll、HQ456312、HQ456313。将9株广州株EV71与EV71的A、B3、B4、B5、cl、c2、C3、c4型及阜阳株全基因组核苷酸序列进行Clustal W比较，发现与C4a型阜阳株等同源性为98％～99％，与C4b同源性为91％一93％，与C1一C3同源性为82％～83％，与B3一B5同源性为81％～83％，与A型同源性为80％。将9株广州分离株EV71的VPl基因与EV71病毒A、B、C型进行ClustalW比较，同样是与C4a为98％～99％，与C4b同源性为92％～94％。与cl。C3的同源性为88％～89％，与Bl～B5型同源性为83％一84％，与A型同源性为81％～82％。将9株广州株与EV71的A、B、C型vPl基因氨基酸序列进行ClnstalW比对，发现VPl基因22位点的谷氨酰胺转变为组氨酸(Q—H)，聚合蛋白中213位点(S—T)和1764位点(V—I)也发生了氨基酸的点突变，P的213位点属于VP2基因，1764位点属于3D基因。结论2008年和2010年分离的9株广州株EV71属于C4a型，与阜阳株同源性为98％～99％，vPl基因22位点发生了点突变．聚合蛋白中213位点和1764位点也发生了氨基酸的点突变。

English Abstract:

Objective To study the genomic genotypes and variation ofhuman enterovirus 71(EV71)infected infants in Guangzhou city。in 2008 and 2010．Methods Primers were designed on the basis of the genomic sequence of EV71 SHZH03 strain(AY465356)in the GenBank，and EV71 genome amplified bv R，r-PCR．PCR-products were directly sequenced and the genomic nucleotide sequences were analyzed with the programs of Clustal W／X．DNASTAR and MEGA 4．1．Results 9 strains of EV71 genome appeared to be 7405 bp in length．The genomic sequences of EV7l Guangzhou strains were compared with those of EV71 iIl GenBank．which revealed that the homology with EV71 genotype C4a Fuyang strains ranged between 98％-99％．Homology with genotype C4b were 92％一94％。with genotypes C1，C2，C3 as 82％一83％，with genotypes B3，B4，B5 as 8l％一83％ and the homology with genotype A was 80％．When compared the VP l genes of EV71 Guangzhou strains with genotypes A，B，C virus，we revealed that the highest homology was also with genotypeC4a．When compared the VPl amino acid sequences of EV7l Guangzhou strains with genotype A，B，Cvirus by Clustal W program．the results revealed t11at the amino acid residue Q at position 22 in VPl gene was transformed to H。while 213(S-+T)and 1764(V_+I)mutations in polyprotein were discovered．Conclusion Data from the sequences and phylogenetics analysis on those Guangzhou strains in 2008 and 2010 revealed that those isolates belong to genotype C4a．with the homology with Fuyang strains as 98％一99％．Mutation of amino acid residue H at position 22 iIl VPl gene was discovered and the neutralizing antibody ofEV7l might have been conversed by this residue．213(S→T)and 1764(V→I)mutations in polyorotein were also discovered．

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