Electrokinetic control was used to prepare a solid-phase nucleic acid hybridization assay in a glass-PDMS microfluidic chip. Streptavidin coated quantum dots (QDs) were immobilized on the derivatized glass surface and subsequently conjugated with probe oligonucleotides. Injection of target nucleic acid and reporter oligonucleotides tagged with Alexa 647 resulted in FRET sensitized photoluminescence from the dye. Measurement of the spatial profiles of FRET sensitized Alexa 647 photoluminescence from the microchannels enabled quantitative detection of femtomole quantities of targets that were 100 basepairs in length. Furthermore, we present how the selectivity of the assay can be tuned by changing QD size and control over oligonucleotide probe density has enabled discrimination against single base-pair mismatches with contrast ratios exceeding two orders of magnitude. The ultimate goal of this work is to provide a lab-on-a-chip diagnostic assay for on demand transduction of pathogens.

Label-free detection of nucleic acid hybridization in an electrokinetically controlled biochip using quantum dots as donors in fluorescence resonance energy transfer

248th National Meeting (2014)

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