Paper-based diagnostic assays are gaining increasing popularity for the development of low cost and easy to use analytical devices that can potentially be employed in remote and resource-limited settings. This presentation will describe the integration of a multiplexed solid-phase quantum dot - fluorescence resonance energy transfer (QD-FRET) nucleic acid hybridization assay on a paper-based platform. The surface of paper was modified with imidazole groups for the assembly of a biorecognition interface comprised of immobilized green-emitting QDs (gQDs) and red-emitting QDs (rQDs) as FRET donors and two types of oligonucleotide probe sequences as biorecognition elements. Addition and subsequent hybridization of Cy3-labeled and Alexa Fluor 647 (A647) oligonucleotide targets provided the proximity for FRET sensitized emission from the acceptor dyes (Cy3 and A647). The gQD/Cy3 FRET pair served as an internal control, while the rQD/A647 FRET pair served to produce the analytical signal for quantitative transduction of nucleic acid hybridization. The limit of detection of the assay in the multicolor format was found to be 90 fmol. Single nucleotide polymorphism discrimination with a contrast ratio of 50 to 1 was possible by using a combination of ionic strength and formamide to control the stringency of nucleic acid hybridization.

Multiplexed transduction of nucleic acid hybridization on a paper-based platform using immobilized quantum dots as donors in fluorescence resonance energy transfer

248th National Meeting (2014)

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