title: A Rapid, Amplification-Free, and Sensitive Diagnostic Assay for Single-Step Multiplexed Fluorescence Detection of MicroRNA.
abstract: The importance of microRNA (miRNA) dysregulation for the development and progression of diseases and the discovery of stable miRNAs in peripheral blood have made these short-sequence nucleic acids next-generation biomarkers. Here we present a fully homogeneous multiplexed miRNA FRET assay that combines careful biophotonic design with various RNA hybridization and ligation steps. The single-step, single-temperature, and amplification-free assay provides a unique combination of performance parameters compared to state-of-the-art miRNA detection technologies. Precise multiplexed quantification of miRNA-20a, -20b, and -21 at concentrations between 0.05 and 0.5 nM in a single 150 μL sample and detection limits between 0.2 and 0.9 nM in 7.5 μL serum samples demonstrate the feasibility of both high-throughput and point-of-care clinical diagnostics.
authors.affiliation: NanoBioPhotonics (nanofret.com), Institut d'Electronique Fondamentale, Université Paris-Sud and CNRS, 91405 Orsay Cedex (France).
authors.forename: Zongwen
authors.initials: Z
authors.lastname: Jin
authors.affiliation: BAM, Federal Institute for Materials Research and Testing, Division 1.10 Biophotonics, Berlin-Adlershof (Germany).
authors.forename: Daniel
authors.initials: D
authors.lastname: Geißler
authors.affiliation: NanoBioPhotonics (nanofret.com), Institut d'Electronique Fondamentale, Université Paris-Sud and CNRS, 91405 Orsay Cedex (France).
authors.forename: Xue
authors.initials: X
authors.lastname: Qiu
authors.affiliation: NanoBioPhotonics (nanofret.com), Institut d'Electronique Fondamentale, Université Paris-Sud and CNRS, 91405 Orsay Cedex (France).
authors.forename: K David
authors.initials: KD
authors.lastname: Wegner
authors.affiliation: NanoBioPhotonics (nanofret.com), Institut d'Electronique Fondamentale, Université Paris-Sud and CNRS, 91405 Orsay Cedex (France). niko.hildebrandt@u-psud.fr.
authors.forename: Niko
authors.initials: N
authors.lastname: Hildebrandt
mesh_terms: D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer; D035683:MicroRNAs / Q000032:analysis*
chemical_list: D035683:MicroRNAs
keywords: FRET; clinical diagnostics; microRNA; multiplexing; time-gated fluorescence detection