Development of a multiplex AmpliDet RNA assay for simultaneous detection and typing of potato virus Y isolates

M. Szemes, M.M. Klerks, J.F.J.M. van den Heuvel, C.D. Schoen

    Research output: Contribution to journalArticleAcademicpeer-review

    21 Citations (Scopus)

    Abstract

    A multiplex AmpliDet RNA assay was developed for the specific detection of potato virus Y (PVY), and for the differentiation of the PVY(N), PVY(O/C) strains and the tuber necrotic isolates (PVY(NTN)). The assay is based on the generic amplification of a region within the coat protein coding region of all known PVY isolates by nucleic acid sequence-based amplification (NASBA) and strain-specific detection by molecular beacons. PVY(NTN)-specific diagnosis is achieved by detecting PVY(N) and PVY(O)-specific sequences flanking a recombination site that is associated with the tuber necrotic pathotype. The assay exhibited good specificity toward the various PVY strains in both single and mixed infections. The technique was validated by the use of 47 PVY isolates originating from six countries. The results of the AmpliDet RNA assay were identical or consistent with those of biological characterisation in the decisive majority of cases
    Original languageEnglish
    Pages (from-to)83-96
    JournalJournal of Virological Methods
    Volume100
    Issue number1-2
    DOIs
    Publication statusPublished - 2002

    Keywords

    • necrotic ringspot disease
    • molecular beacons
    • amplification
    • pvyntn
    • nasba
    • necrosis
    • sequence
    • genome
    • probes
    • strain

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