J Biomed Nanotechnol. 2020 Feb 1;16(2):166-178. doi: 10.1166/jbn.2020.2889.
White root rot (WRR) disease caused by , a fungal pathogen, results in severe damage to various fruit trees, decreasing their marketability. Regular monitoring is a major process because the pathogen can remain in the soil around the host for a long time. Loop-mediated isothermal amplification (LAMP) is a highly sensitive and efficient amplification technology of nucleic acids (DNA or RNA) that can be performed at constant temperatures. Thus, it has been spotlighted as a useful tool for detecting several infectious agents. In the present study, LAMP-based Turn-on Fluorescent Paper (ToFP) devices were designed and applied to detect . LAMP conditions were optimized and found to be optimal at a reaction temperature (62 °C) and a reaction time (30 minutes). These reaction conditions were confirmed by applying them to infectious soil samples collected from the field. The limitation of detection was identified as 10 fg of genomic DNA under optimized LAMP conditions. These LAMP-based ToFP devices were generated with easily available stationery materials and the utility of these devices to analyze the LAMP results were confirmed through several experiments on a total of 14 field samples. The results showed that the developed LAMP-based detection system was very sensitive and had the advantages of rapid detection and high availability in the field.
白根腐烂病（WRR）是由一种真菌病原体引起的，会对各种果树造成严重损害，降低其市场价值。定期监测是一个主要过程，因为病原体可以在宿主周围的土壤中长时间存活。环介导等温扩增（LAMP）是一种高度敏感和高效的核酸（DNA 或 RNA）扩增技术，可以在恒温下进行。因此，它已被视为检测几种传染病原体的有用工具。在本研究中，设计了基于 LAMP 的荧光开启纸（ToFP）设备，并应用于检测 。优化了 LAMP 条件，发现反应温度（62°C）和反应时间（30 分钟）为最佳条件。通过应用于从田间采集的感染性土壤样本，证实了这些反应条件。在优化的 LAMP 条件下，检测的极限为 10 fg 基因组 DNA。这些基于 LAMP 的 ToFP 设备是用易得的办公用品制作的，并通过对总共 14 个田间样本的多项实验，证实了这些设备在分析 LAMP 结果方面的实用性。结果表明，开发的基于 LAMP 的检测系统非常灵敏，具有快速检测和现场高可用性的优点。
