Direct detection of potato leafroll virus in potato tubers by immunocapture and the isothermal nucleic acid amplification method NASBA

G. Leone, H.B. van Schijndel, B. van Gemen, C.D. Schoen

    Research output: Contribution to journalArticleAcademicpeer-review

    25 Citations (Scopus)

    Abstract

    NASBA, an isothermal amplification method for nucleic acids, was applied to the detection of RNA of potato leafroll virus (PLRV) in a single enzymatic reaction at 41 °C. A set of primers was selected from the coat protein open reading frame sequence of PLRV to allow amplification of viral RNA. The NASBA reaction products were visualized after electrophoresis by ethidium bromide or acridine orange staining. The specificity of the amplification products was validated by Northern blot analysis with a PLRV-specific 32P-labelled oligonucleotide probe. The procedure was coupled to immunocapture of PLRV virions from tuber extracts by immobilized antibodies in microtubes. It was possible to discriminate readily by this method between uninfected and primarily PLRV-infected potato tubers. NASBA is suitable for the direct detection of PLRV in potato tubers from primarily infected plants, offering the potential to considerably simplify the inspection of seed-potatoes for virus infection.
    Original languageEnglish
    Pages (from-to)19-27
    JournalJournal of Virological Methods
    Volume66
    DOIs
    Publication statusPublished - 1997

    Fingerprint Dive into the research topics of 'Direct detection of potato leafroll virus in potato tubers by immunocapture and the isothermal nucleic acid amplification method NASBA'. Together they form a unique fingerprint.

  • Cite this