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

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.