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Rapid diagnostic methods for Phytophthora tentaculata using LAMP
Pages: 1-6
Year: Issue:  6
Journal: Journal of Nanjing Forestry University(Natural Science Edition)

Keyword:  loop-mediated isothermal amplification ( LAMP )hydroxynaphthol blue;
Abstract: In order to identify the disease caused by Phytophthora tentaculata, and keep the spread of the disease under control and reduce the economic losses quickly and accurately, loop?mediated isothermal amplification ( LAMP ) assay targeting the PtYpt1 element was used to detect Phytophthora tentaculata. The PtYpt1-LAMP assay amplified the target element in less than 80 min at 64℃ efficiently and was evaluated specificitily and sensitivitily. The specificity was eval?uated against Phytophthora tentaculata, Phytophthora spp., Pythium spp., and true fungi isolates. The P. tentaculata DNA products were visualised as a ladder?like banding pattern by 2% gel electrophoresis. A positive colour ( sky blue) was only observed in the presence of P. tentaculata by addition of hydroxynaphthol blue prior to amplification, whereas none of other isolates showed a colour change. The detection limit of the PtYpt1-specific LAMP assay for P. tentaculata was 1 ng/μL of genomic DNA per reaction.
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