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Visual Detection of Potato leafroll virus by One‐step Reverse Transcription Loop‐Mediated Isothermal Amplification of DNA with Hydroxynaphthol Blue Dye
Author(s) -
Ahmadi Sayvan,
Almasi Mohammad A.,
Fatehi Foad,
Struik Paul C.,
Moradi Aboubakr
Publication year - 2013
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/jph.12037
Subject(s) - loop mediated isothermal amplification , reverse transcription loop mediated isothermal amplification , potato leafroll virus , biology , microbiology and biotechnology , agarose , dna , reverse transcriptase , virology , agarose gel electrophoresis , virus , gene , rna , plant virus , biochemistry
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 ( 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 .