Chines Journal of Vector Biology and Control ›› 2013, Vol. 24 ›› Issue (5): 392-396.DOI: 10.11853/j.issn.1003.4692.2013.05.004

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Determination and analysis of complete genomic sequences of three Dengue 2 virus strains isolated in Guangdong province, China

BAI Zhi-jun1, ZHANG Pei2, HONG Wen-yan2, LIU Jian-wei2, DI Biao1, YANG Zhi-cong1, REN Rui-wen2, FANG Mei-yu2, LIN Li-hui2   

  1. 1 Guangzhou Center for Disease Control and Prevention, Guangzhou 510080, Guangdong Province, China;
    2 Center for Disease Control and Prevention of Guangzhou Military District, Guangzhou 510507, Guangdong Province, China
  • Received:2013-04-26 Online:2013-10-20 Published:2013-10-20
  • Supported by:

    Supports by the National Institutes of Health Project (No. 1R01AI083202-01A1), National Science and Technology Major Project of China (No. 2012ZX10004-213-005), Guangdong Science and Technology Department Program (No. 2011B031500011), Guangzhou Science and Technology Department Program (No. 2010Y1-C151, 2012Y2-00020) and Science and Technology Program of Guangzhou Health Department (No. 201102A212006, 20121A011122)

广东省流行的3株登革热2型病毒基因组全序列测定及分析

白志军1, 张培2, 洪文艳2, 刘建伟2, 狄飚1, 杨智聪1, 任瑞文2, 方美玉2, 林立辉2   

  1. 1 广州市疾病预防控制中心病毒免疫科, 广东 广州 510080;
    2 广州军区疾病预防控制中心, 广东 广州 510507
  • 通讯作者: 任瑞文, Email: renruiwen@hotmail.com
  • 作者简介:白志军(1977- ),男,硕士,副主任技师,主要从事病原学研究。Email: junewhite@163.com;张培(1979- ),女,硕士,助理研究员,主要从事病原学研究。Email: zhangpei7982@163.com
  • 基金资助:

    美国国立卫生研究院项目(1R01AI083202-01A1); 国家科技重大专项(2012ZX10004-213-005); 广东省科技计划项目(2011B031500011); 广州市科技计划项目(2010Y1-C151,2012Y2-00020); 广州市医药卫生科技项目(201102A212006, 20121A011122)

Abstract:

Objective To determine the complete genomic sequences of 3 Dengue 2 virus (DEN-2) strains isolated in Guangdong province, China and to investigate their genotypes and sources. Methods The complete genomic sequences of 3 DEN-2 strains (GD09/93, GD05/98, and GD19/2001) from Guangdong province were established by RT-PCR amplification. The phylogenetic tree of DEN-2 was constructed by Kimura method. Results The complete genomic sequences of 3 DEN-2 strains were 10 723 nucleotides (nt) in length and contained a single long open reading frame (ORF) of 10 173 nucleotides (97-10 269 nt), encoding 3391 amino acids. The ORF encoded structural and non-structural proteins, flanked by 5' and 3' non-coding regions. Comparing GD05/98 with GD09/93, GD05/98 with GD19/2001, and GD09/93 with GD19/2001, the nucleotide sequence homologies were 93.3%, 92.4%, and 97.6%, respectively, and the deduced amino acid sequence homologies were 96.7%, 96.5%, and 98.5%, respectively. Conclusion The DEN-2 strains are pathogenic in suckling mice. Compared with DEN-2-04 strain (nonpathogenic in suckling mice), there are 18 amino acid substitutions that confer changes in charge or polarity. The charge changes at PrM-134, NS2A-153, and NS4B-102 have a relatively strong impact on the antigenicity. GD05/98 strain falls within the groupⅡ comprising Thailand strains, while GD09/93 and GD19/2001 strains fall within the group Ⅳ comprising strains from Indonesia, Australian and Taiwan. There are different DEN genotypes in China, and one genotype of DEN-2 may be transmitted in different periods.

Key words: Dengue virus, Complete genomic sequence, Phylogenetic tree, Genotype, Polymerase chain reaction

摘要:

目的 测定广东省3株登革热2型病毒的全序列,探讨其来源及基因型。方法 运用RT-PCR法扩增来自广东省的登革热2型GD09/93、GD05/98和GD19/2001病毒株的全序列,采用遗传距离法构建进化树。结果 3株登革热2型病毒的全基因组长度均为10 723 nt, 5'及3'端各有一非编码区,结构基因和非结构基因位于基因组97~10 269 nt之间,共编码3391个氨基酸,读码结构完全相同。GD05/98与GD09/93、GD05/98与GD19/2001、GD09/93与GD19/2001之间的碱基序列同源性分别为93.3%、92.4%和97.6%,氨基酸序列同源性分别为96.7%、96.5%和98.5%。结论 3株登革热2型病毒均对乳鼠致病,与对乳鼠不致病的参考株(DEN2-04株)比较共有18个氨基酸位点的差异引起极性或电荷变化,PrM-134、NS2A-153、NS4B-102所带电荷的变化对抗原性影响较大。GD05/98株与泰国分离株同为Ⅱ基因型,GD09/93和GD19/2001株与印度尼西亚、澳大利亚、台湾株分在Ⅳ基因型; 表明我国登革热2型病毒有不同的基因型,而不同时期也存在同一种基因型传播。

关键词: 登革热病毒, 全序列, 系统发生树, 基因型, 聚合酶链反应

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