NTT Hi-Tech Institute, Nguyen Tat Thanh University, Ho Chi Minh City, Vietnam.
College of Veterinary Medicine, Vietnam National University of Agriculture, Hanoi, Vietnam.
Transbound Emerg Dis. 2021 Jul;68(4):2595-2602. doi: 10.1111/tbed.13879. Epub 2020 Nov 4.
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.
非洲猪瘟（ASF）是一种具有高传染性和高死亡率的病毒性疾病。越南最近一次的 ASF 疫情始于 2019 年，对周边亚洲国家构成了传播威胁。由于没有商业疫苗或有效的化学疗法，因此需要快速诊断和必要的生物安全措施来控制该疾病。尽管世界动物卫生组织推荐的 ASF 诊断方法是实时 PCR，但包括主混合物设置、模板提取和同时测试多个样本的高成本 qPCR 设备在内的理想诊断程序并不便携。在这项研究中，修改并评估了一种用于检测 ASF 病毒的比色环介导的等温扩增（LAMP）检测方法，该方法使用了 2019 年越南国内猪只采集的粗血清样本。LAMP 结果可在 30 分钟内通过肉眼观察到，无需提取 DNA 和使用复杂的设备。直接比色 LAMP 检测的灵敏度、特异性和检测限与商业诊断实时 PCR 试剂盒相当。结果强烈表明，适应的比色 LAMP 检测法具有用于 ASF 现场诊断的巨大潜力。
