title: 5-color multiplexed microwave-accelerated metal-enhanced fluorescence: detection and analysis of multiple DNA sequences from within one sample well within a few seconds.
abstract: We present a potentially highly sensitive and selective bio-assay for the potential detection of any five different DNA sequences from one sample in one well. The assay is based on a DNA "rapid catch and signal" (DNA-RCS) technology developed for the detection of different DNA sequences from a sample well area. Our signal amplification utilizes the metal-enhanced fluorescence (MEF) of dyes attached to the probe-DNAs, which hybridizes with the pre-formed mixture of anchor-DNA scaffolds on silver island films (SiFs). Low-power microwave irradiation accelerates both the formation of the anchor-DNA scaffold on the SiF-surface and anchor/probe DNA hybridization, i.e. "rapid catch" of target DNAs from a bulk solution, decreasing the assay run time from hours to only a few seconds. Localization of signaling dye-labels close to the SiFs make them extremely photostable, which allows for collecting/integrating the signal over a long time period. To demonstrate a 5 color DNA assay (5-plex) we have used a range of readily available Alexa™ dyes. Advantages and perspectives of the RCS-technologies ability to detect 5 different DNA sequences from within one plate-well are discussed.
authors.affiliation: Institute of Fluorescence and Department of Chemistry and Biochemistry, UMBC, 701 East Pratt Street, Baltimore, MD, 21202, USA.
authors.forename: Anatoliy
authors.initials: A
authors.lastname: Dragan
authors.forename: Chris D
authors.initials: CD
authors.lastname: Geddes
mesh_terms: D015342:DNA Probes / Q000737:chemistry*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D008670:Metals / Q000737:chemistry*; D008872:Microwaves; D009693:Nucleic Acid Hybridization; D017422:Sequence Analysis, DNA / Q000379:methods*; D012834:Silver / Q000737:chemistry*
chemical_list: D015342:DNA Probes; D005456:Fluorescent Dyes; D008670:Metals; D012834:Silver