Development of rapid in-field loop-mediated isothermal amplification (LAMP) assays for phytoplasmas

Loop-mediated isothermal amplification (LAMP) is an isothermal amplification technique that can be undertaken with minimal equipment to obtain amplification of target DNA within 30 minutes. A range of assays for specific 16Sr phytoplasma groups, which when combined with rapid DNA extraction techniques can result in detection of the phytoplasma in plant material within 1 hour of sampling was developed. A range of alternative methods are available for detection of the amplification product including incorporation of hydroxyl napthol blue into the reagent mix, agarose gel electrophoresis and real-time detection systems. The advantage of the real-time method is that tubes do not have to be opened, minimising the risk of contamination of samples.