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

作者：Zongwen Jin, Daniel Geißler, Xue Qiu, K. David Wegner, Niko Hildebrandt · 发表于：Angewandte Chemie International Edition · 年份：2015 · DOI：10.1002/anie.201504887 · 被引用次数：189 · 研究领域：Advanced biosensing and bioanalysis techniques、MicroRNA in disease regulation、RNA Interference and Gene Delivery

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.