Development of a multiplex AmpliDet RNA for the simultaneous detection of Potato leafroll virus and Potato virus Y in potato tubers

M.M. Klerks, G.O.M. Leone, M. Verbeek, J.F.J.M. van den Heuvel, C.D. Schoen

    Research output: Contribution to journalArticleAcademicpeer-review

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    Abstract

    A novel isothermal multiplex AmpliDet RNA system is described for the simultaneous amplification and detection of Potato leafroll virus (PLRV) and Potato virus Y (PVY) in seed potatoes. The risk of contamination by carry-over during diagnostic screening is eliminated by performing the reaction in a single closed tube. The viruses present in a sample are identified using differently coloured molecular beacons directed to a selected virus-specific sequence within the amplicon formed during amplification. With this system, as little as 10 fg of purified PLRV or PVY can be detected. The presence of both viruses in a sample is detected by the multiplex assay within a high range of virus concentrations. The reliability of the multiplex assay was compared with the enzyme-linked immunosorbent assay for detection of PLRV- or PVY-antigens in potato tubers. The multiplex assay detected clearly the viruses present originally in the potato tubers in all samples, demonstrating its potential for routine diagnostic work and high-throughput screening.
    Original languageEnglish
    Pages (from-to)115-125
    JournalJournal of Virological Methods
    Volume93
    Issue number1-2
    DOIs
    Publication statusPublished - 2001

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    Potyvirus
    Solanum tuberosum
    RNA
    Viruses
    Seeds
    Enzyme-Linked Immunosorbent Assay
    Antigens

    Cite this

    Klerks, M.M. ; Leone, G.O.M. ; Verbeek, M. ; van den Heuvel, J.F.J.M. ; Schoen, C.D. / Development of a multiplex AmpliDet RNA for the simultaneous detection of Potato leafroll virus and Potato virus Y in potato tubers. In: Journal of Virological Methods. 2001 ; Vol. 93, No. 1-2. pp. 115-125.
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    abstract = "A novel isothermal multiplex AmpliDet RNA system is described for the simultaneous amplification and detection of Potato leafroll virus (PLRV) and Potato virus Y (PVY) in seed potatoes. The risk of contamination by carry-over during diagnostic screening is eliminated by performing the reaction in a single closed tube. The viruses present in a sample are identified using differently coloured molecular beacons directed to a selected virus-specific sequence within the amplicon formed during amplification. With this system, as little as 10 fg of purified PLRV or PVY can be detected. The presence of both viruses in a sample is detected by the multiplex assay within a high range of virus concentrations. The reliability of the multiplex assay was compared with the enzyme-linked immunosorbent assay for detection of PLRV- or PVY-antigens in potato tubers. The multiplex assay detected clearly the viruses present originally in the potato tubers in all samples, demonstrating its potential for routine diagnostic work and high-throughput screening.",
    author = "M.M. Klerks and G.O.M. Leone and M. Verbeek and {van den Heuvel}, J.F.J.M. and C.D. Schoen",
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    Development of a multiplex AmpliDet RNA for the simultaneous detection of Potato leafroll virus and Potato virus Y in potato tubers. / Klerks, M.M.; Leone, G.O.M.; Verbeek, M.; van den Heuvel, J.F.J.M.; Schoen, C.D.

    In: Journal of Virological Methods, Vol. 93, No. 1-2, 2001, p. 115-125.

    Research output: Contribution to journalArticleAcademicpeer-review

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    AU - Klerks, M.M.

    AU - Leone, G.O.M.

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    AU - van den Heuvel, J.F.J.M.

    AU - Schoen, C.D.

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    AB - A novel isothermal multiplex AmpliDet RNA system is described for the simultaneous amplification and detection of Potato leafroll virus (PLRV) and Potato virus Y (PVY) in seed potatoes. The risk of contamination by carry-over during diagnostic screening is eliminated by performing the reaction in a single closed tube. The viruses present in a sample are identified using differently coloured molecular beacons directed to a selected virus-specific sequence within the amplicon formed during amplification. With this system, as little as 10 fg of purified PLRV or PVY can be detected. The presence of both viruses in a sample is detected by the multiplex assay within a high range of virus concentrations. The reliability of the multiplex assay was compared with the enzyme-linked immunosorbent assay for detection of PLRV- or PVY-antigens in potato tubers. The multiplex assay detected clearly the viruses present originally in the potato tubers in all samples, demonstrating its potential for routine diagnostic work and high-throughput screening.

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