4.2 Article

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

Journal

JOURNAL OF PHYTOPATHOLOGY
Volume 161, Issue 2, Pages 120-124

Publisher

WILEY
DOI: 10.1111/jph.12037

Keywords

Potato leafroll virus; one-step RT-LAMP; hydroxynaphthol blue dye

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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.

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