Detection of oligonucleotides is important in diagnosis and treatment of various cancers and genetic disorders. A variety of methods are available for detection of nucleic acids that are dependent on hybridization for signal generation. Fluorescence based detection is one such method. While fluorescence based methods offer good sensitivity, the need for target labeling makes sample preparation more time consuming. Labeling of the target can be avoided by means of incorporating an intercalating dye such as a derivative of a thiazole orange into the analytical system by dual labeling of the probe molecule. The use of such dyes has been previously reported. The fluorescence intensity of these dyes is dependent on the interaction with the DNA duplex. In the absence of the target the two dye molecules form an H-aggregate which allows for excitonic interactions that lead to quenching of fluorescence. Hybridization of the target molecule results in dissociation of the aggregate and regeneration of fluorescence emission. An assay has been developed where DNA probe molecules that are double labeled with thiazole orange derivative are excited by fluorescent resonance energy transfer (FRET) from quantum dot (QD) donors. Immobilizing the dually labeled probe molecules on the surface of the QDs averts the need for labeling of target, allows for signal enhancement due to one-donor-multiple acceptor interactions, and provides practical advantages such as ratiometric detection.

Towards solid-phase assays using intrinsically labeled oligonucleotide probes on quantum dots for<br />determination of selective hybridization

249th National Meeting (2015)

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