The Food and Environment Research Agency, Sand Hutton, York, UK.
Lett Appl Microbiol. 2010 Dec;51(6):650-7. doi: 10.1111/j.1472-765X.2010.02949.x. Epub 2010 Oct 12.
To develop a sensitive, rapid and simple method for detection of Botrytis cinerea based on loop-mediated isothermal amplification (LAMP) that would be suitable for use outside a conventional laboratory setting.
A LAMP assay was designed based on the intergenic spacer of the B. cinerea nuclear ribosomal DNA (rDNA). The resulting assay was characterized in terms of sensitivity and specificity using DNA extracted from cultures. The assay consistently amplified 65 pg B. cinerea DNA. No cross-reactivity was observed with a range of other fungal pathogens, with the exception of the closely related species Botrytis pelargonii. Use of a novel real-time LAMP platform (the OptiGene Genie I) allowed detection of B. cinerea in infected rose petals, with amplification occurring in <15 min.
The LAMP assay that was developed is suitable for rapid detection of B. cinerea in infected plant material.
The LAMP method combines the sensitivity and specificity of nucleic acid-based methods with simplified equipment and a reduced reaction time. These features make the method potentially suitable for on-site use, where the results of testing could help to inform decisions regarding the storage and processing of commodities affected by B. cinerea, such as cut flowers, fruit and vegetables.
开发一种基于环介导等温扩增（LAMP）的敏感、快速且简单的检测方法，以用于常规实验室以外的环境。
基于灰葡萄孢核核糖体 DNA（rDNA）的基因间间隔区设计了 LAMP 检测方法。使用从培养物中提取的 DNA 对该检测方法的灵敏度和特异性进行了特征描述。该检测方法可一致地扩增 65 pg 的灰葡萄孢 DNA。除了密切相关的物种 Pelargonii botrytis 外，该检测方法与一系列其他真菌病原体无交叉反应。新型实时 LAMP 平台（OptiGene Genie I）的使用允许在受感染的玫瑰花瓣中检测到灰葡萄孢，扩增在<15 分钟内发生。
所开发的 LAMP 检测方法适用于快速检测感染植物材料中的灰葡萄孢。
LAMP 方法将基于核酸的方法的灵敏度和特异性与简化的设备和缩短的反应时间相结合。这些特点使该方法具有潜在的现场使用价值，测试结果可帮助确定受灰葡萄孢影响的商品（如切花、水果和蔬菜）的储存和处理方式。
