肠道病毒71型景德镇分离株全基因组序列分析

摘要

Objective To analyze the complete genome of enterovirus 7l( EV71 )strain ( 2011JDZ35 )isolated in Jing-dezhen( 2011 ). Methods The virus was isolated from clinical samples including throat swab from patients with severe hand-foot-mouth disease( HFMD ). The EV71 strain was identified by reverse transcript PCR( RT-PCR ),and the isolated EV71 whole genomes were amplified by RT-PCR with seven specific primer pairs and were compared with EV71 A,B,C and CA16( G10 ), which were available from GenBank by homogeneity and phylogenetic tree analyses. Results The full genomic length of 2011JDZ35 was 7406 bp, which contained 5'UTR ( 742 bp ),the open reading frame ( ORF,6582 bp ) and 3'UTR( 82 bp ). The ORF encoded a polyprotein with 2193 amino acid residues. It was found that 2011JDZ35 was similar to FY08-C30-P14, EV71/Ningbo. CHN/065/2010 and HZ08/Hangzhou/2008 by means of sequence analysis. The nucleotide and amino acid sequence identity was 97.3%, 97.3% and 97.7% , and 99. 1%,99. 2%,99. 2%,respectively, belonging to C4 subgenotype. 2011JDZ35 shared lower homology with Cox. A16 and BrCr. The nucleotide and amino acid sequence identity was 77.5%,80.0% and 89. 8% ,94. 8%, respectively. Conclusion The genome structure of 2011JDZ35 strain is consistent with enterovirus. The phylogenetic relationships of the 2011JDZ35 strain with other EV71 strains are different. Ningbo strain is the nearest to 2011JDZ35 strain, and Fuyang and Hangzhou strains are nearer to 2011JDZ35 strain. The 2011JDZ35 strain is different from Malaysia, Taiwan and Xiamen strains, and dramatically different from the standard strain. 2011JDZ35 and Ningbo strains probably originate from the same EV71 strain, which is one of the prevalent strains in mainland China in 2008.%目的 分析2011年景德镇地区重症手足口病患儿标本肠道病毒71型(EV71)分离株2011JDZ35全基因组序列特征.方法 采集重症手足口病患儿咽拭子标本,进行病毒分离和逆转录-聚合酶链反应(RT-PCR)扩增分离病毒的全基因组序列,并进行全基因组核苷酸序列测定,参考EV71 A、B、C各基因型的参考毒株和国内外分离毒株进行同源性分析并构建系统发生树.结果 2011JDZ35株的基因组长度为7406 bp,其中包括5′端非编码区(5′UTR)长742 bp,病毒基因组编码区(ORF)全长6582 bp及3′端非编码区(3′UTR)长82 bp.ORF编码含2193个氨基酸残基的多聚蛋白.2011JDZ35株基因组的结构与FY08-C30-P14、EV71/Ningbo.CHN/065/2010、HZ08/Hangzhou/2008十分接近,整个基因组的核苷酸同源性分别为97.3%,97.3%,97.7%;氨基酸同源性为99.1%,99.2%,99.2%.均属于C4亚型,而与Cox.A16国际标准株G10(U05876)及EV71国际标准株BrCr(U22521)差异较大,核苷酸同源性分别仅为77.5%,80.0%;氨基酸同源性为89.8%,94.8%.结论 EV71型病毒2011JDZ35株全基因组的组成和结构符合肠道病毒特征,与其他EV71型病毒具有相同的基因组结构,2011JDZ35株与中国大陆浙江宁波株亲缘关系最近,与阜阳株和杭州株等中国大陆内陆地区分离株较近,而与马来西亚、我国台湾、厦门分离株的差异较为明显,与国际标准株则有较大差异;2011JDZ35株与浙江宁波株具有共同的进化途径,推断2011JDZ35株与浙江宁波株可能共同来源于2008年中国大陆地区流行的EV71病毒株同一种系.