Angew Chem Int Ed Engl. 2015 Aug 17;54(34):10024-9. doi: 10.1002/anie.201504887. Epub 2015 Jul 29.

A Rapid, Amplification-Free, and Sensitive Diagnostic Assay for Single-Step Multiplexed Fluorescence Detection of MicroRNA.

Angewandte Chemie (International ed. in English)

Zongwen Jin, Daniel Geißler, Xue Qiu, K David Wegner, Niko Hildebrandt

PMID: 26226913 DOI: 10.1002/anie.201504887

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.

© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Keywords: FRET; clinical diagnostics; microRNA; multiplexing; time-gated fluorescence detection

Substances

• MicroRNAs

MeSH terms

• Fluorescence*
• Fluorescence Resonance Energy Transfer*
• MicroRNAs / analysis

Publication Types

• Journal Article
• Research Support, Non-U.S. Gov't