[2008年在北京分离的肠道病毒71型全基因组的遗传特征分析]

[Genetic characterization of enterovirus 71 complete genome isolated in Beijing, 2008].

作者信息

Yao Xin, Mao Qun-Ying, Huang Wei-Jin, He Peng, Zhou Cheng, Zhang Wei, Liang Zheng-Lun, Li Feng-Xiang, Wang Jun-Zhi

机构信息

National Institute for Control of Pharmaceutical and Biological Products, Beijing, China.

出版信息

Zhonghua Liu Xing Bing Xue Za Zhi. 2009 Jul;30(7):729-32.

PMID:19957602

Abstract

OBJECTIVE

To investigate the characterization of the complete genome of EV71 in Beijing, 2008 and to provide basis for selecting appropriate virus strain to develop vaccine.

METHODS

12 throat swab samples were collected from children with hand-foot-mouth disease (HFMD). One sample named 08YM-3 was cultured and isolated in vero cells. Viral RNA was extracted and carried out by RT-PCR and 5', 3' rapid amplification of cDNA ends (RACE) to obtain the sequence from 08YM-3. PCR products were cloned and analyzed. Nucleotide identity between sequences was calculated and sequence alignments were made to generate phylogenetic trees using MegAlign in DNAStar.

RESULTS

3 clones were constructed that covered EV71 complete genome. Data from sequences analysis showed that this viral strain named BJ08 shared 95.6%-96.7%, 88.3%-96.1%, 78.1%-94.0%, 90.8%-94.6%, 85.9%-94.1% and 90.9%-93.9% in 5' UTR, P1, P2, P3, 3' UTR region and complete genome with C4 subtype, respectively. BJ08 showed low nucleotides identity (<90%) with other subtypes. Phylogenetic trees established from alignment of the complete genome and VP1 region indicated that BJ08 belonged to C4 subtype. BJ08 and C4 subtype strains shared the same amino acids in 6 sites in VP1 region, which were associated with EV71 subtype. There was no mutation in VP1 antigen epitope (92-107aa).

CONCLUSION

This BJ08 strain belonged to C4 subtype. Further study on EV71 complete genome would have great significance for vaccine research.

摘要

目的

研究2008年北京肠道病毒71型（EV71）全基因组特征，为筛选合适毒株研发疫苗提供依据。

方法

采集12例手足口病（HFMD）患儿咽拭子标本。将其中一份名为08YM - 3的标本在vero细胞中培养分离。提取病毒RNA，通过逆转录聚合酶链反应（RT - PCR）及5′、3′端cDNA末端快速扩增（RACE）获得08YM - 3的序列。对PCR产物进行克隆及分析。计算序列间核苷酸同一性，使用DNAStar中的MegAlign进行序列比对并构建系统发育树。

结果

构建了覆盖EV71全基因组的3个克隆。序列分析数据显示，该毒株BJ08在5′非编码区（UTR）、P1、P2、P3、3′UTR区域及全基因组与C4亚型的核苷酸同一性分别为95.6% - 96.7%、88.3% - 96.1%、78.1% - 94.0%、90.8% - 94.6%、85.9% - 94.1%和90.9% - 93.9%。BJ08与其他亚型的核苷酸同一性较低（<90%）。基于全基因组和VP1区域比对构建的系统发育树表明BJ08属于C4亚型。BJ08与C4亚型毒株在VP1区域6个位点的氨基酸相同，这些位点与EV71亚型相关。VP1抗原表位（92 - 107aa）无突变。