Silva Gonçalo, Boonham Neil
Natural Resources Institute, University of Greenwich, Chatham Maritime, UK.
School of Natural and Environmental Sciences, Agriculture Building, Newcastle University, Newcastle upon Tyne, UK.
Methods Mol Biol. 2025;2912:157-164. doi: 10.1007/978-1-0716-4454-6_13.
A real-time loop-mediated isothermal amplification (LAMP) is described to diagnose cassava mosaic disease (CMD). The disease continues to cause great damage in Sub-Saharan African countries threatening food security. Early detection of the pathogen is paramount to control the spread of the disease and promote the safe exchange of clean planting material. Real-time LAMP in combination with a crude but rapid method of DNA extraction can be used to detect the two most widely distributed species of cassava mosaic geminiviruses in Sub-Saharan Africa, African cassava mosaic virus (ACMV) and East African cassava mosaic virus (EACMV), in less than 30 min at a single operation temperature (65 °C) without the need for any expensive instrumentation. Further, this protocol avoids the laborious extraction of high-quality DNA and making it suitable to be used in poorly resourced laboratories and in field settings.
本文描述了一种用于诊断木薯花叶病(CMD)的实时环介导等温扩增(LAMP)方法。该疾病继续在撒哈拉以南非洲国家造成巨大破坏,威胁粮食安全。尽早检测到病原体对于控制疾病传播和促进清洁种植材料的安全交换至关重要。实时LAMP结合一种粗糙但快速的DNA提取方法,可用于检测撒哈拉以南非洲两种分布最广泛的木薯花叶双生病毒,即非洲木薯花叶病毒(ACMV)和东非木薯花叶病毒(EACMV),在单一操作温度(65°C)下不到30分钟即可完成,无需任何昂贵的仪器设备。此外,该方案避免了费力地提取高质量DNA,使其适用于资源匮乏的实验室和野外环境。