Digital Mirna Detection Based on Target Cycling-Induced Fret Signal Amplification

Absolute quantitative analysis of microRNA (miRNA) plays a vital role in disease diagnosis and cancer studies. Conventional digital miRNA detection methods usually used cDNA obtained from reverse transcription PCR of miRNA as templates, which quantified miRNA indirectly and suffered from low accuracy. This paper reports a novel miRNA detection method based on target cycling-induced FRET signal amplification and droplet microfluidics. It allows, for the first time, direct digital miRNA detection with high selectivity and sensitivity at single-molecule level theoretically. In this method, the full use of miRNA sequence information made the hybridization between targets and DNA probes unique and sensitive, offering high capacity to distinguish single-base mismatches.