2010—2011年湖南省郴州市手足口病EV71型VP1区基因序列分析  

Sequence analysis of VP1 gene of human enterovirus 71 in Chenzhou City,Hunan Province,2010 - 2011

作　　者：郑文[1] 谭徽[1] 谢群[1] 陈白鹭[1] 陈柏塘[1] 刘爱平[1] 

机构地区：[1]湖南省郴州市疾病预防控制中心,湖南郴州423000

出　　处：《华南预防医学》2013年第3期7-11,共5页South China Journal of Preventive Medicine

摘　　要：目的了解2010—2011年湖南省郴州市手足口病EV71病毒的基因特征,为手足口病的预防控制提供科学依据。方法从2010—2011年郴州市各县(市、区)送检的手足口病肛拭子标本中随机挑选60份标本采用RD细胞进行病毒分离,采用Real time-RT-PCR方法检测肠道病毒核酸,挑选14株致细胞病变阳性且EV71核酸检测阳性的标本(其中2010年的4株均为郴州市手足口流行夏季高峰期分离株,2011年的毒株主要为冬季高峰期分离株),采用RT-PCR法扩增出EV71分离株的VP1区片段,随后进行VP1区核苷酸序列测定和分析,并使用生物信息学方法作基因特性分析。结果郴州市14株EV71毒株在生物进化树上与C4a亚型的代表株属同一分支。14株EV71毒株之间核苷酸序列的同源性在96.1%～100.0%之间,氨基酸序列的同源性在99.3%～100.0%之间;与A、B、C各基因型和亚型代表株EV71 VP1编码区核苷酸和氨基酸序列同源性分析表明,郴州各分离株与安徽阜阳2008年毒株C4a亚型代表株(EU703812)的同源性最高,其中核苷酸同源性在91.8%～93.3%之间,氨基酸同源性均为99.3%。14株分离株与安徽阜阳2008年毒株(C4a亚型代表株)VP1区段的氨基酸编码序列进行比较,发现K98E、S283T和A293S 3处位点变异,重症病例与普通病例的EV71病毒VP1氨基酸变异无明显差异。结论 2010—2011年郴州市手足口病的优势毒株EV71属于C4a基因亚型。郴州市各县(市、区)的EV71亲缘关系近,毒株基因较稳定,未发现氨基酸突变位点与病例类型有关联。Objective To study the genetic characterization of human enterovirus 71 （EV -71 ） in Chenzhou City, Hunan Province during 2010 to 2011, and provide scientific data for HFMD control and prevention. Methods 60 anal swab specimens sent by each county （city, district） of Chenzhou were se- lected randomly for virus isolation. Reahime RT-PCR was used for detecting enterovirus RNA. Specimens of 14 strains （Four strains of them were isolated at the summer epidemic peak in 2010, and the others were mainly isolated at the winter epidemic peak in 2011 ） with cytopathic and EV71 RNA positive were ampli- fied by RT-PCR, and then nucleotide sequencing was performed and a phylogenetic tree was constructed u- sing bioinformatics method. Results 14 EVT1 strains were clustered with CAa subgenotype reference strains （EU703812） of C genotype in phylogenetic tree. The 14 selected strains showed high homology （96. 1% - 100.0% and 99. 3% - 100.0% ） in nucleotide and translated nucleotide homology analysis, and higher nucleotide homology （91.8% -93.3% ） and amino acid homology （99. 3% ） with the Fuyang isolated strains （CAa subgenotype reference strain, EU703812） than A, B, and C subgenotype reference strains. K98E, S283T, and A293S were found between Chenzhou isolated strains and Fuyang isolated strains （EU703812）. The significant difference of nucleotide or amino acid sequence was not found be- tween HEV71 strains from severe （dead） patients and mild patients. Conclusion EV71 was the predominant pathogen that caused HFMD in Chenzhou City in 2010 and 2011, which belonged to C4a genotype. High homology and stability were found in the 14 isolated EV71 strains of Chenzhou. No amino acid muta- tions were found to be associated with the case type.

关 键 词：手足口病 人肠道病毒71型 基因 

分 类 号：R512.5[医药卫生—内科学]