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Abstract

This invention relates to a bioassay method for detecting and/or quantitating a short single-stranded nucleic acid analyte employing a binary probe system, where at least one of the two discrete oligonucleotide probe parts of the binary probe has partially double-stranded (self-complementary) stem-loop structure at one terminus and single-stranded overhang sequence region at the other terminus, where the single-stranded terminal regions of both discrete parts of the binary probe hybridize to adjacent complementary regions in the sequence of the nucleic acid analyte molecule, and at least one discrete part of the binary probe comprising a stem-loop structure and single-stranded overhang sequence region hybridizes to terminal region in the sequence of the nucleic acid analyte molecule forming a nick structure. The binary probe system employed in the bioassay method is based on a luminescent reporter technology, either lanthanide chelate complementation or resonance energy transfer with lanthanide label as a donor. Thereby the method allows detection and/or quantitation of the short nucleic acid analyte molecule by time-resolved fluorometry.

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First Claim