Department of Plant Pathology, Washington State University, Pullman, WA, United States of America.
Agdia, Inc., Elkhart, IN, United States of America.
PLoS One. 2022 Aug 1;17(8):e0270918. doi: 10.1371/journal.pone.0270918. eCollection 2022.
Potato mop-top virus (PMTV) is considered an emerging threat to potato production in the United States. PMTV is transmitted by a soil-borne protist, Spongospora subterranean. Rapid, accurate, and sensitive detection of PMTV in leaves and tubers is an essential component in PMTV management program. A rapid test that can be adapted to in-field, on-site testing with minimal sample manipulation could help in ensuring the sanitary status of the produce in situations such as certification programs and shipping point inspections. Toward that goal, a rapid and highly sensitive recombinase polymerase amplification (RPA)-based test was developed for PMTV detection in potato tubers. The test combines the convenience of RPA assay with a simple sample extraction procedure, making it amenable to rapid on-site diagnosis of PMTV. Furthermore, the assay was duplexed with a plant internal control to monitor sample extraction and RPA reaction performance. The method described could detect as little as 10 fg of PMTV RNA transcript in various potato tissues, the diagnostic limit of detection (LOQ) similar to that of traditional molecular methods.
马铃薯帚顶病毒（PMTV）被认为是美国马铃薯生产的一种新出现的威胁。PMTV 通过土壤传播的原生动物——地下泡囊海绵菌传播。快速、准确、灵敏地检测叶片和块茎中的 PMTV 是 PMTV 管理计划的重要组成部分。一种可以适应现场、现场测试的快速检测方法，只需最小限度的样本处理，有助于在认证计划和发货点检查等情况下确保农产品的卫生状况。为此，开发了一种基于重组酶聚合酶扩增（RPA）的快速、高灵敏度检测马铃薯块茎中 PMTV 的方法。该测试将 RPA 检测的便利性与简单的样本提取程序相结合，使其能够快速进行现场诊断 PMTV。此外，该检测方法与植物内参进行了双重检测，以监测样本提取和 RPA 反应性能。所描述的方法可以检测到各种马铃薯组织中低至 10 fg 的 PMTV RNA 转录本，检测限（LOD）与传统分子方法相似。
