Rapid Detection of Phytophthora ramorum and P. kernoviae by Two-Minute DNA Extraction Followed by Isothermal Amplification and Amplicon Detection by Generic Lateral Flow Device

A method for nucleic-acid-based detection of pathogens in plant material has been developed which comprises a Simple and rapid method for extracting DNA oil the nitrocellulose membranes of lateral-flow devices. loop-mediated Isothermal amplification (LAMP) of target DNA using labeled primers, and detection of the generically labeled amplification products by a Sandwich immunoassay in a lateral-flow-device format Each of these steps can be performed without specialist equipment and is suitable for on-site use, and a result can be obtained in just over an hour. A LAMP assay for the detection of plant DNA (cytochrome oxidase gene) call be used in conjunction with pathogen-specific assays to confirm negative results The use of this method is demonstrated for file detection of Phytophthora ramorum the causal agent of sudden oak death and dieback/leaf blight in a range of tree. shrub. and herbaceous species. and the recently described pathogen P kernoviae