Molecular beacon probes combined with amplification by NASBA enable homogeneous, real-time detection of RNA

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

    Research output: Contribution to journalArticleAcademicpeer-review

    240 Citations (Scopus)

    Abstract

    Molecular beacon probes can be employed in a NASBA amplicon detection system to generate a specific fluorescent signal concomitantly with amplification. A molecular beacon, designed to hybridize within the target sequence, was introduced into NASBA reactions that amplify the genomic RNA of potato leafroll virus (PLRV). During amplification, the probe anneals to the antisense RNA amplicon generated by NASBA, producing a specific fluorescent signal that can be monitored in real-time. The assay is rapid, sensitive and specific. As RNA amplification and detection can be carried out in unopened vessels, it minimizes the risk of carry-over contaminations. Robustness has been verified on real-world samples. This homogeneous assay, called AmpliDet RNA, is a significant improvement over current detection methods for NASBA amplicons and is suitable for one-tube applications ranging from high-throughput diagnostics to in vivo studies of biological activities.
    Original languageEnglish
    Pages (from-to)2150-2155
    JournalNucleic acids research
    Volume26
    Issue number9
    DOIs
    Publication statusPublished - 1998

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