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 ( F 3, B 3, 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 .