Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, Sichuan, PR China.
Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, Sichuan, PR China.
J Virol Methods. 2020 Nov;285:113885. doi: 10.1016/j.jviromet.2020.113885. Epub 2020 May 31.
African swine fever (ASF) is an acute, hemorrhagic, highly contagious disease caused by African swine fever virus (ASFV) infection of domestic pigs and wild boars, showing mortality rates up to 100 %. There are no effective vaccines or antiviral drugs available for ASFV. Therefore, disease control is mainly based on animal slaughtering and the enforcement of strict sanitary measures. In order to establish a rapid, sensitive and simple method for on-site detection of ASFV, a recombinase polymerase amplification (RPA) combined with lateral flow dipstick (LFD) was developed using a pair of specific primers and probe. Using recombinant plasmid pMD19-T-K205R DNA as a template, the RPA-LFD detection could be accomplished in 10 min at a temperature of 36℃-44℃. More specific than PCR and more rapid and simpler than real-time PCR, RPA-LFD has the same detection limit of 1 × 10 copies/reaction as real-time PCR, also with no cross-reaction with other viral strains. A convenient and rapid ASFV RPA-LFD detection method was developed, which will provide an efficient method for investigating epidemiology of ASFV infection.
非洲猪瘟（ASF）是一种由非洲猪瘟病毒（ASFV）感染家猪和野猪引起的急性、出血性、高度传染性疾病，死亡率高达 100%。目前尚无有效的 ASF 疫苗或抗病毒药物。因此，ASF 的防控主要基于对病猪和感染猪的扑杀，并执行严格的卫生措施。为了建立一种快速、灵敏、简便的现场检测 ASF 的方法，本研究使用一对特异性引物和探针，开发了一种重组酶聚合酶扩增（RPA）与侧向流动试纸条（LFD）相结合的检测方法。以重组质粒 pMD19-T-K205R DNA 为模板，在 36℃-44℃的温度下，RPA-LFD 检测可在 10 分钟内完成。与 PCR 相比，RPA-LFD 具有更高的特异性，与实时 PCR 相比，具有更快、更简单的优点，其检测限与实时 PCR 相同，为 1×10 拷贝/反应，与其他病毒株也无交叉反应。本研究建立了一种简便、快速的 ASF RPA-LFD 检测方法，为 ASF 感染的流行病学调查提供了一种有效的方法。
