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Volume 36 Issue 2
April 2021

    Lei Zuo, Zengxu Song, Yi Zhang, Xiwen Zhai, Yaru Zhai, Xueran Mei, Xin Yang and Hongning Wang. Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick for On-Site Diagnosis of African Swine Fever Virus[J]. Virologica Sinica, 2021, 36(2): 325-328. doi: 10.1007/s12250-020-00309-z
    Citation: Lei Zuo, Zengxu Song, Yi Zhang, Xiwen Zhai, Yaru Zhai, Xueran Mei, Xin Yang, Hongning Wang. Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick for On-Site Diagnosis of African Swine Fever Virus .VIROLOGICA SINICA, 2021, 36(2) : 325-328.  http://dx.doi.org/10.1007/s12250-020-00309-z
    shu

    环介导等温扩增结合横向流动试纸条现场检测非洲猪瘟病毒的研究

    cstr: 32224.14.s12250-020-00309-z
    • 1.

      四川大学生命科学学院，动物疫病防控与食品安全四川省重点实验室生物，资源与生态环境教育部重点实验室，成都 610064

    • 2.

      四川省动物疫病预防控制中心，成都 610041

    • 通讯作者： 王红宁, whongning@163.com, ORCID: http://orcid.org/0000-0002-7244-9929
    • 收稿日期： 2020-07-31
      录用日期： 2020-09-18
      出版日期： 2020-11-06

    • 由非洲猪瘟病毒（African swine fever virus，ASFV）导致的非洲猪瘟病（African swine fever，ASF）最高可以造成100%死亡，严重威胁世界养猪业。截止目前，还没有有效的ASF疫苗。因此，快速、灵敏、适用于现场检测的方法是目前控制ASFV传播所急需的。本研究结合环介导等温扩增（loop-mediated isothermal amplification，LAMP）与横向流动试纸条（lateral flow dipstick，LFD），针对ASFV中B646L基因保守序列，建立了一种新的ASFV的LAMP-LFD检测方法，并通过与靶向同一区域的PCR、巢式PCR、qPCR及电泳检视的LAMP方法对比，对其特异性和灵敏性进行了研究。特异性试验表明，本研究建立的所有方法都具有ASFV特异性，不会与其他常见猪病毒发生交叉反应。灵敏性试验表明，LAMP方法（电泳检视的LAMP和LAMP-LFD）在几种方法中是最灵敏的，检测限为100.6B646L片段每反应。在模拟临床样品检测试验中，PCR、巢式PCR、qPCR检测限均降低了10倍，而LAMP方法的检测限无变化，表明LAMP方法更耐受组织样品中非靶标DNA的干扰。另外，在对52份临床样品的检测中，LAMP方法仍表现最高阳性检出率（16/52）, 而PCR、巢式PCR及qPCR的阳性检出率分别为13/52、14/52和15/52。综上所述，本研究建立的灵敏、特异的LAMP-LFD方法可应用于ASFV感染的临床诊断和现场监测。

    Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick for On-Site Diagnosis of African Swine Fever Virus

    • 1.

      College of Life Sciences, Sichuan University, Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, Key Laboratory of Bio-Resource and EcoEnvironment of Ministry of Education, Chengdu 610064, China

    • 2.

      Sichuan Provincial Center for Animal Disease Control and Prevention, Chengdu 610041, China

    • Corresponding author: Hongning Wang, whongning@163.com
    • ORCID: http://orcid.org/0000-0002-7244-9929
    • Received Date: 31 July 2020
      Accepted Date: 18 September 2020
      Published Date: 06 November 2020
    • 加载中

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      15. Wang DG, Yu JH, Wang YZ, Zhang M, Li P, Liu M, Liu YH (2020) Development of a real-time loop-mediated isothermal amplification (LAMP) assay and visual LAMP assay for detection of African swine fever virus (ASFV). J Virol Methods 276:10

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    • 加载中

    • Electronic Supplementary Material

      10.1007s12250-020-00309-z-ESM.pdf

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      Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick for On-Site Diagnosis of African Swine Fever Virus

        Corresponding author: Hongning Wang, whongning@163.com
      • 1. College of Life Sciences, Sichuan University, Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, Key Laboratory of Bio-Resource and EcoEnvironment of Ministry of Education, Chengdu 610064, China
      • 2. Sichuan Provincial Center for Animal Disease Control and Prevention, Chengdu 610041, China
      • Received Date: 31 July 2020
      • Accepted Date: 18 September 2020
      • Published Date: 06 November 2020

      Abstract: 

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