SPECTRALLY-RESOLVED CHEMILUMINESCENT PROBES FOR SENSITIVE MULTIPLEX MOLECULAR QUANTIFICATION

摘要

Hybrid luminescent probes emit light of distinct wavelength ranges and intensities upon energy transfer from an in-common, acridinium ester chemiluminophore to a coupled luminophore. The probes include: (1) a target binding region with a base sequence that is substantially complementary to a portion of the target nucleic acid sequence; (2) an acridinium ester (AE) moiety attached to a first region flanking the target binding region; (3) a luminophore coupled to the AE moiety to allow energy transfer from an acridone moiety, produced by a chemical triggering of the AE moiety, to the luminophore; and (4) a quencher moiety attached to a second region flanking the target binding region, such that the first and second flanking regions are on the opposite sides of the target binding region. The probes are particularly useful in homogeneous assays for sensitive, multiplex quantification of nucleic acid target sequences without prior enzymatic amplification.