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A Rapid, Amplification‐Free, and Sensitive Diagnostic Assay for Single‐Step Multiplexed Fluorescence Detection of MicroRNA
Author(s) -
Jin Zongwen,
Geißler Daniel,
Qiu Xue,
Wegner K. David,
Hildebrandt Niko
Publication year - 2015
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201504887
Subject(s) - microrna , multiplexing , förster resonance energy transfer , multiplex , computational biology , nucleic acid , point of care , microbiology and biotechnology , fluorescence , biology , bioinformatics , computer science , medicine , gene , genetics , physics , pathology , telecommunications , quantum mechanics
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 n M in a single 150 μL sample and detection limits between 0.2 and 0.9 n M in 7.5 μL serum samples demonstrate the feasibility of both high‐throughput and point‐of‐care clinical diagnostics.