一株引起病毒性脑炎的埃可病毒30型分离株的VP1基因序列特征分析  

Analysis of characters of VP1 gene sequence of Echovirus 30 A363/KM/2009 strain isolated from patients with viral meningitis

作　　者：陈俊英[1] 潘玥[1] 吉玛[1] 张云昆[1] 朱云[1] 朱艳菊[1] 邵聪文[1] 马绍辉[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所云南省重大传染病疫苗研发重点实验室,云南昆明650118

出　　处：《中国生物制品学杂志》2014年第2期163-167,共5页Chinese Journal of Biologicals

基　　金：云南省应用基础研究项目(2013FZ136)

摘　　要：目的分析2009年昆明市病毒性脑炎患儿粪便中分离获得的埃可病毒30型（Echovirus30,Echo30）分离株A363/KM/2009全长VP1基因序列特征。方法从疑似病毒性脑膜炎患儿粪便样本中分离Echo30,提取病毒RNA,采用RT-PCR法扩增病毒VP1基因,并进行测序。采用NCBI BLAST软件对所测定的序列进行数据库比对;采用Geneious软件对Echo30分离株全长VP1基因的核苷酸及氨基酸同源性进行分析;采用Mega 5.1软件对A363/KM/2009分离株进行VP1基因分析,并与20个参考株的VP1序列进行比较。结果分离株为A363/KM/2009,其VP1基因的核苷酸长度与其他Echo30一致,均为867 bp;与其他Echo30分离株核苷酸同源性在76.9%-95.7%之间,氨基酸同源性在88.7%-99.7%之间;与中国株的核苷酸和氨基酸的同源性分别为84.5%-95.7%和96.9%-99.7%,其中与中国浙江分离株Echo30/zhejiang/10/4的同源性最高,为95.7%;与广西分离株GX10/15的同源性最低,为84.5%。氨基酸序列分析显示,与其他中国株包括脑脊液分离株相比,未见特异的突变位点;与国外分离株的核苷酸和氨基酸同源性分别为76.9%-86.3%和88.7%-97.3%;基因进化树分析显示,与其他中国株分属不同的两个分支,而与Echo30/zhejiang/10/4和Echo30/zhejiang/4/04（CSF）株亲缘关系较近。结论 A363/KM/2009分离株为肠道病毒Echo30,属于中国两个分支中的一支。Objective To analyze the genetic characters of full-length VP1 gene sequence of Echovirus 30（Echo30） A363 / KM / 2009 isolated from fecal specimens of children with viral meningitis in Kunming City,Yunnan Province, China in 2009. Methods Echo30 was isolated from the fecal specimens of children with viral meningitis,from which vi- ral RNA was extracted,and VP1 gene was amplified by RT-PCR and sequenced. The sequencing result was compared with those in data bank by using NCBI BLAST software. The homologies of nucleotide and amino acid sequences of full- length VP1 gene of the isolate were analyzed by using Geneious software. The VP1 gene sequence of the isolate was com- pared to those of 20 reference strains by using Mega 5. 1 software. Results The nucleotide length of isolated A363 / KM / 2009 strain was 867 bp,which was consistent with those of other Echo30 strains. The homologies of nucleotide and amino acid of the isolate were 76. 9% - 95. 7% and 88. 7% - 99. 7% to those of other Echo30 isolates,and 84. 5% - 95. 7% and 96. 9% - 99. 7% to those of isolates in other provinces in China,respectively. The homology of nucleotide of A363 / KM / 2009 to that of Echo30 / Zhejiang / 10 / 14（95. 7%）was the highest,while that to GX10 / 15（84. 5%）was the lowest. Amino sequencing showed no specific mutation site of A363 / KM / 2009 compared with other Chinese isolates including those isolated from cerebrospinal fluid. The homologies of nucleotide and amino acid of A363 / KM / 2009 were 76. 9% - 86. 3% and 88. 7% - 97. 3% to those of other isolates abroad respectively. Phylogenetic analysis revealed that the sequences of Chinese strains segregated into two distinct clusters. However,the evolutional relationship of A363 / KM / 2009 strain was the closest to Echo30 / zhejiang / 10 / 4（CSF）strain. Conclusion A363 / KM / 2009 strain was Echo30 belonging to one of the two clusters in China.

关 键 词：病毒性脑炎 埃可病毒30型 VP1基因 序列分析 

分 类 号：R373.24[医药卫生—病原生物学] Q78[医药卫生—基础医学]