Visual Detection of Potato leafroll virus by One-step Reverse Transcription Loop-Mediated Isothermal Amplification of DNA with Hydroxynaphthol Blue Dye

Abstract

Loop-mediated isothermal amplification (LAMP) assay is a novel technique for amplifying DNA under constant temperature, with high specificity, sensitivity, rapidity and efficiency. We applied reverse transcription loop-mediated isothermal amplification (RT-LAMP) to visually detect Potato leafroll virus. One-step RT-LAMP was performed using RNA of PLRV-infected potato leaves and a set of primers (F3, B3, FIP, BIP, LF and LB) designed for RT-LAMP reaction of the coat protein (CP) gene of PLRV. Positive effects of RT-LAMP were detected by agarose gel electrophoresis and hydroxynaphthol blue (HNB) dye and were shown by a colour change from violet to sky blue. RT-LAMP with HNB dye proved to be a simple assay for the rapid detection of PLRV. © 2012 Blackwell Verlag GmbH.