Rapid and Multiplexed MicroRNA Diagnostic Assay Using Quantum Dot-Based Förster Resonance Energy Transfer

• Published in: ACS nano Vol. 9; no. 8; pp. 8449 - 8457
• Main Authors: Qiu, Xue, Hildebrandt, Niko
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 25.08.2015
• Subjects: Assaying; Avidin - chemistry; Biological Assay; Chemical Sciences; Diagnostic systems; Energy transfer; Fluorescence Resonance Energy Transfer; Fretting; Humans; Life Sciences; Limit of Detection; Luminescence; MicroRNAs - blood; Multiplexing; Oligonucleotides - chemistry; Physics; Quantum dots; Quantum Dots - chemistry; Ribonucleic acids; Semiconductors; Solutions; Streptavidin - chemistry; Terbium - chemistry; Time Factors; terbium; miRNA; FRET; clinical diagnostics; multiplexing; nanocrystals
• ISSN: 1936-0851; 1936-086X
• DOI: 10.1021/acsnano.5b03364

The detection of next generation microRNA (miRNA) biomarkers has become a highly important aspect for clinical diagnostics. We use multiplexed Förster resonance energy transfer (FRET) between a luminescent Tb complex and three different semiconductor quantum dots (QDs) to sensitively detect three different miRNAs from a single 150 μL sample with ca. 1 nM (subpicomol) detection limits. The rapid and amplification-free mix-and-measure assay format is based on careful design of miRNA base pairing and stacking to selectively detect different miRNAs with very strong sequence homologies. Clinical applicability is demonstrated by sensitive multiplexed quantification of three miRNAs at low (2 to 10 nM) and varying concentrations in samples that contained up to 10% serum.