Direct colorimetric LAMP assay for rapid detection of African swine fever virus: A validation study during an outbreak in Vietnam
Direct colorimetric LAMP assay for rapid detection of African swine fever virus: A validation study during an outbreak in Vietnam
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DOI：
10.1111/tbed.13879
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发表时间：
2020-11-04
影响因子：
4.3
通讯作者：
Van Phan Le
中科院分区：
文献类型：
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作者：
Diem Hong Tran;Hau Thi Tran;Van Phan Le
非洲猪瘟（ASF）是一种具有高致死率的高传染性病毒性疾病。越南最近一次非洲猪瘟疫情始于2019年，有可能蔓延到亚洲邻国。由于没有商业化的疫苗或有效的化疗药物，需要快速诊断和必要的生物安全程序来控制这种疾病。虽然世界动物卫生组织推荐的非洲猪瘟诊断方法是实时PCR，但理想的诊断程序包括主混合物设置、模板提取和高成本的qPCR设备，无法同时检测许多样品。本研究对比色环介导等温扩增（LAMP）方法进行了改进，并对该方法在2019年疫情期间从越南家猪采集的粗血清样本中检测非洲猪瘟病毒进行了评估。LAMP结果可在30分钟内肉眼可见，不需要DNA提取和复杂的设备。直接比色LAMP法的灵敏度、特异性和检测限与商业实时诊断PCR试剂盒相当。结果表明，适应比色LAMP法在ASF现场诊断中具有显著的潜力。
African swine fever (ASF) is a highly infectious viral disease with high mortality. The most recent ASF outbreak in Vietnam began in 2019, posing a threat to spread to the neighbouring Asian countries. Without a commercial vaccine or efficient chemotherapeutics, rapid diagnosis and necessary biosecurity procedures are required to control the disease. While the diagnostic method of ASF recommended by the World Organization of Animal Health is real-time PCR, the ideal diagnosis procedure including master mix setup, template extraction and a high-cost qPCR equipment for many samples being tested simultaneously is not portable. In this study, a colorimetric loop-mediated isothermal amplification (LAMP) assay was modified and evaluated for ASF virus detection using crude serum samples collected from domestic pigs in Vietnam during the 2019 outbreak. The LAMP results can be readily visualized to the naked eye within 30 min without the requirement of DNA extraction and sophisticated equipment. The sensitivity, specificity and limit of detection of direct colorimetric LAMP assay were comparable to a commercial diagnostic real-time PCR kit. Results strongly indicate that the adapted colorimetric LAMP assay has a remarkable potential for the in-field diagnosis of ASF.