| 摘要 |
<p>Detection probes enabling very convenient and highly accurate and sensitive detection of DNAs or RNAs having specific base sequences in specimens in the presence of the probes in excess of the target nucleic acid; and a detection method. When mixed with a specimen containing the target substance (DNA, RNA, etc.) having a polynucleotide having a specific base sequence, two types of fluorescent label detection probes hybridize, while being adjacent to each other, with the target nucleic acid. As a result, there arises a shift of the resonance energy between the two types of fluorescent dye molecules. When the resonance energy shift occurs in the hybrid thus formed, the fluorescence attenuation of the acceptor fluorescent dye molecule can be sufficiently delayed compared with the fluorescence attenuation of the directly excited acceptor by appropriately regulating (1) the number of bases between the two nucleotides to which the fluorescent dye moleculees have bonded; (2) the structure (double- or single-stranded) of the hybrid between the two nucleotides to which the fluorescent dye molecules have bonded; and (3), on the detection probes, the sites of the nucleotides into which the fluorescent dye molecules are to be introduced; and by (4) using a pair of detection probes with the appropriate selection of the fluorescent dye molecules. The detection method comprises using the above-mentioned detection probes and detecting the target substance by measuring changes in the fluorescence attenuation curve of the acceptor after the irradiation with pulse excitation rays, thus enabling highly accurate and sensitive detection of the target nucleic acid in the presence of the detection probes in excess of the target nucleic acid.</p> |
