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    朱俊灵, 叶佐东, 邓洁汝, 勾红潮, 陈金顶. 猪瘟病毒RT-LAMP-LFD检测方法的建立与应用[J]. 华南农业大学学报, 2016, 37(1): 1-7. DOI: 10.7671/j.issn.1001-411X.2016.01.001
    引用本文: 朱俊灵, 叶佐东, 邓洁汝, 勾红潮, 陈金顶. 猪瘟病毒RT-LAMP-LFD检测方法的建立与应用[J]. 华南农业大学学报, 2016, 37(1): 1-7. DOI: 10.7671/j.issn.1001-411X.2016.01.001
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    ZHU Junling, YE Zuodong, DENG Jieru, GOU Hongchao, CHEN Jinding. Rapid detection of classical swine fever virus by loop-mediatedisothermal amplification combined with lateral flow dipstick method[J]. Journal of South China Agricultural University, 2016, 37(1): 1-7. DOI: 10.7671/j.issn.1001-411X.2016.01.001
    Citation: ZHU Junling, YE Zuodong, DENG Jieru, GOU Hongchao, CHEN Jinding. Rapid detection of classical swine fever virus by loop-mediatedisothermal amplification combined with lateral flow dipstick method[J]. Journal of South China Agricultural University, 2016, 37(1): 1-7. DOI: 10.7671/j.issn.1001-411X.2016.01.001
    shu

    猪瘟病毒RT-LAMP-LFD检测方法的建立与应用

    Rapid detection of classical swine fever virus by loop-mediatedisothermal amplification combined with lateral flow dipstick method

      摘要
    • 摘要:
      目的 建立一种新型的、能在基层场地进行实时便捷诊断的猪瘟病毒检测方法。
      方法 环介导等温扩增技术(Loop-mediated isothermal amplification, LAMP)-横向流动试纸条技术(Lateral flow dipstick, LFD)相结合，针对猪瘟病毒(Classical swine fever virus, CSFV) NS5B基因设计一套特异性引物及生物素标记探针，结合异硫氰酸荧光素标记的扩增产物，建立RT-LAMP-LFD检测方法。
      结果 所建立的方法能够特异地检测出CSFV的存在，检测PRRSV、JEV、PCV-2等其他病毒均为阴性；所建立的CSFV-RT-LAMP方法对RNA的最低检测限为50 pg；反应快速，整个扩增反应可在1 h内完成；操作简单, 不需要PCR仪等复杂的仪器，结果可经肉眼观察。
      结论 应用RT-LAMP-LFD检测CSFV特异性强、灵敏度高，并且操作安全、简便、快捷，可以满足基层检疫的需要。

       

      Abstract:
      Objective The experiment was conducted to establish a rapid real time detection method of classical swine fever virus at local fields.
      Method A rapid detection method(RT-LAMP-LFD) of classical swine fever virus(CSFV) was developed by loop-mediated isothermal amplification (LAMP) combined with lateral flow dipstick (LFD) method. The LAMP method for amplification of the CSFV NS5B gene using fluorescein isothiocyanate-labeled primer was combined with a chromatographic LFD for rapid and simple visual detection of CSFV.
      Result This method could specifically detect CSFV, but not other virus such as PRRSV, JEV and PCV-2. The minimum detection limit of the CSFV-RT-LAMP-LFD method was 50 pg. The detection could be finished within 1 h without using complex instruments such as PCR machine and the results were visible by eye.
      Conclusion This RT-LAMP-LFD method is a highly specific and sensitive, safe, rapid and simple way for field detection of CSFV infection.

       

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