Thiazole orange derivatives can be used in designing bioassays for detection of nucleic acids based on an “on-off” switching system. One such assay has been published where a DNA probe was doubly labeled with thiazole orange derivatives. In the absence of the fully complementary (FC) target the dyes form an H-aggregate which results in quenching of fluorescence emission due to excitonic interactions between the dyes. The hybridization of the FC target to the probe causes dissociation of the aggregate as the dyes intercalate into the double stranded duplex. This results in the restoration of fluorescence emission. A new approach is proposed where the doubly labeled probes are immobilized on the surface of Quantum Dots (QDs). The QDs act as donors in Fluorescence Resonance Energy Transfer (FRET) used to excite the dyes. This configuration reduces the need for labeling the target, provides with improved selectivity as well as practical advantages such as ratiometric detection. The signal from the dye was optimized in terms of the type of linkage used to attach the dyes to the probe, location of the dye within the probe as well as the distance between the dyes. The optimized system was then assessed for quantitative response and selectivity.

Towards assays using intrinsically labeled oligonucleotide probes on QDs for detection of selective hybridization

251st National Meeting (2016)

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