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Volume 24 Issue 1
February 2009

    Xian QI, Yong-jun JIAO, Hao PAN, Lun-biao CUI, Wei-xing FAN, Bao-xu HUANG, Zhi-yang SHI and Hua WANG. Genetic Analysis and Rescue of a Triple-reassortant H3N2 Influenza A Virus Isolated From Swine in Eastern China *[J]. Virologica Sinica, 2009, 24(1): 52-58. doi: 10.1007/s12250-009-3006-7
    Citation: Xian QI, Yong-jun JIAO, Hao PAN, Lun-biao CUI, Wei-xing FAN, Bao-xu HUANG, Zhi-yang SHI, Hua WANG. Genetic Analysis and Rescue of a Triple-reassortant H3N2 Influenza A Virus Isolated From Swine in Eastern China * .VIROLOGICA SINICA, 2009, 24(1) : 52-58.  http://dx.doi.org/10.1007/s12250-009-3006-7
    shu

    一株三源基因重配的H3N2亚型流感病毒的拯救和遗传进化分析*
    Genetic Analysis and Rescue of a Triple-reassortant H3N2 Influenza A Virus Isolated From Swine in Eastern China*

    cstr: 32224.14.s12250-009-3006-7
    • 1.

      江苏省疾病预防控制中心-南京大学联合病毒研究所，江苏省疾病预防控制中心，江苏南京，210009

    • 2.

      .中国动物卫生与流行病学中心，山东青岛，266032

    • 通讯作者： 汪华, hua@jscdc.cn
    • 收稿日期： 2008-10-13
      录用日期： 2008-12-18

    • 从猪体内分离一株H3N2亚型流感病毒(A/Swine/Shangdong/3/2005， Sw/SD/3/2005)。全基因组序列分析表明Sw/SD/3/2005是一株三源基因重配病毒，其中HA, NA, NP, M, PB1和PA基因来源于人H3N2病毒，PB2基因来源于人H1N1病毒，而NS基因则来源于古典猪H1N1病毒。Sw/SD/3/2005的分离进一步为猪是A型流感病毒“基因混合器”观点提供了证据，同时也提示应该加强A型流感病毒在猪体内的生态分布和分子进化的监测研究。将经RT-PCR得到的A/swine/Shandong/3/2005八个基因的cDNA片段分别克隆到含有pol Ⅰ和pol Ⅱ启动子和终止子序列的PHW2000质粒,得到了8 个可在真核细胞中复制和表达的质粒。将这8个质粒共转染293T 细胞,培养6h 后吸取上清液,转接入SPF鸡胚和MDCK细胞中,再经过72h 培养,得到病毒血凝滴度为1：8 。继续接种鸡胚，经过3次传代后，病毒血凝价稳定在1：256。Sw/SD/3/2005的成功拯救为进一步深入研究H3N2亚型流感病毒跨物种传播的分子机制奠定了基础
      Abstract One influenza H3N2 virus, A/swine/Shandong/3/2005 (Sw/SD/3/2005), was isolated from pigs with respiratory disease on a farm in eastern China. Genetic analysis revealed that Sw/SD/3/2005 was a triple-reassortant virus with a PB2 gene from human-like H1N1, NS from classical swine H1N1, and the remaining genes from human-like H3N2 virus. These findings further support the concept that swine can serve as reservoir or mixing vessels of influenza virus strains and maintain genetic and antigenic stability of viruses. Furthermore, we have successfully established a reverse genetics system based on eight plasmids and rescued Sw/SD/3/2005 through cell transfection. HI tests and RT-PCR confirmed that the rescued virus maintained the biological properties of the wild type Sw/SD/3/2005. The successful establishment of the reverse genetics system of Sw/SD/3/2005 will enable us to conduct extensive studies of the molecular evolution of H3N2 influenza viruses in swine.

    Genetic Analysis and Rescue of a Triple-reassortant H3N2 Influenza A Virus Isolated From Swine in Eastern China *

    • 1.

      Jiangsu CDC-Nanjing University Joint Institute of Virology, Jiangsu Center for Disease Control and Pevention, Nanjing 210009, China

    • 2.

      China Animal Health and Epidemiology Center, Qingdao 266032, China

    • Corresponding author: Hua WANG, hua@jscdc.cn
    • Received Date: 13 October 2008
      Accepted Date: 18 December 2008

      Fund Project: China Postdoctoral Science Foundation 20070411041Jiangsu Postdoctoral Science Foun-dation 0701032B

    • One influenza H3N2 virus, A/swine/Shandong/3/2005 (Sw/SD/3/2005), was isolated from pigs with respiratory disease on a farm in eastern China. Genetic analysis revealed that Sw/SD/3/2005 was a triple-reassortant virus with a PB2 gene from human-like H1N1, NS from classical swine H1N1, and the remaining genes from human-like H3N2 virus. These findings further support the concept that swine can serve as reservoir or mixing vessels of influenza virus strains and maintain genetic and antigenic stability of viruses. Furthermore, we have successfully established a reverse genetics system based on eight plasmids and rescued Sw/SD/3/2005 through cell transfection. HI tests and RT-PCR confirmed that the rescued virus maintained the biological properties of the wild type Sw/SD/3/2005. The successful establishment of the reverse genetics system of Sw/SD/3/2005 will enable us to conduct extensive studies of the molecular evolution of H3N2 influenza viruses in swine.
    • 加载中

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      Genetic Analysis and Rescue of a Triple-reassortant H3N2 Influenza A Virus Isolated From Swine in Eastern China *

        Corresponding author: Hua WANG, hua@jscdc.cn
      • 1. Jiangsu CDC-Nanjing University Joint Institute of Virology, Jiangsu Center for Disease Control and Pevention, Nanjing 210009, China
      • 2. China Animal Health and Epidemiology Center, Qingdao 266032, China
      • Received Date: 13 October 2008
      • Accepted Date: 18 December 2008
      Fund Project:  China Postdoctoral Science Foundation 20070411041Jiangsu Postdoctoral Science Foun-dation 0701032B

      Abstract: One influenza H3N2 virus, A/swine/Shandong/3/2005 (Sw/SD/3/2005), was isolated from pigs with respiratory disease on a farm in eastern China. Genetic analysis revealed that Sw/SD/3/2005 was a triple-reassortant virus with a PB2 gene from human-like H1N1, NS from classical swine H1N1, and the remaining genes from human-like H3N2 virus. These findings further support the concept that swine can serve as reservoir or mixing vessels of influenza virus strains and maintain genetic and antigenic stability of viruses. Furthermore, we have successfully established a reverse genetics system based on eight plasmids and rescued Sw/SD/3/2005 through cell transfection. HI tests and RT-PCR confirmed that the rescued virus maintained the biological properties of the wild type Sw/SD/3/2005. The successful establishment of the reverse genetics system of Sw/SD/3/2005 will enable us to conduct extensive studies of the molecular evolution of H3N2 influenza viruses in swine.

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