Multiplex DNA Detection Without Fluorophore Labeling

Dakota Technologies, Inc., Fargo ND

Investigators

Abstract

[unreadable] DESCRIPTION (provided by applicant): A cost-effective and field-deployable DNA detection system is urgently needed to rapidly and sensitively screen crude biological samples for a plethora of pathogens, including biological warfare agents. Dakota Technologies, Inc.'s (DTI) detection approach involves DNA hybridization duplexes that have been stained with a simple DNA staining dye. Although such dyes show very large fluorescence enhancements upon binding to DNA, non-specific binding to the single-strand probes and to single-stranded flanking regions of the target DNA must be overcome. Preliminary data proves DTI's innovative fluorescence lifetime measurement technology yields extremely sensitive, specific, and quantitative discrimination of the dye bound to the hybridized double-stranded region. Not having to use expensive fluorophore-labeled probes or targets offers two very significant advantages: greatly reduced cost and the ability to design probes unencumbered by complicated and highly restrictive rules. Beads that have been labeled with specific DNA probes will be assembled according to a specified order in a capillary whose inner diameter is slightly greater than the bead diameter. The result is highly effective affinity capture in a multiplex detection format. Rapid (<5 min) affinity capture and detection of low quantity (<100 amol) target DNA from multiple analytes in crude sample mixtures will be demonstrated by the end of Phase I. [unreadable] [unreadable] [unreadable] [unreadable]