Double-Strand Displacement Biosensor and Quencher-Free Fluorescence Strategy for Rapid Detection of MicroRNA

Abstract

We describe a facile quencher-free fluorescence strategy for rapid detection of microRNAs (miRNAs) by using a novel double-strand displacement sensor. The sensor is designed with an outstanding 2-aminopurine (2-AP) fluorophore as a probe and a predesigned cDNA, which can completely complement the target miRNA and partly complement the 2-AP probe. When the target miRNA is added, the cDNA can be competed off from the cDNA\2-AP probe duplex, thereby forming a cDNA\RNA heteroduplex. The free 2-AP probe induces an increase in the fluorescent signal. A limit of detection of 5 nM and a wide linear range from 5 to 1000 nM (R(2) = 0.9971) are achieved by this assay. The rapid detection strategy can be accomplished within 2 h without expensive nanoparticles and complicated instruments for the whole procedure, thus, offering a significant potential for clinical application.