Issue 42, 2018

Advanced microRNA-based cancer diagnostics using amplified time-gated FRET


MicroRNAs (miRNAs) play an important role in cellular functions and in the development and progression of cancer. Precise quantification of endogenous miRNAs from different clinical patient and control samples combined with a one-to-one comparison to standard technologies is a challenging but necessary endeavor that is largely neglected by many emerging fluorescence technologies. Here, we present a simple, precise, sensitive, and specific ratiometric assay for absolute quantification of miRNAs. Isothermally amplified time-gated Förster resonance energy transfer (TG-FRET) between Tb donors and dye acceptors resulted in miRNA assays with single-nucleotide variant specificity and detection limits down to 4.2 ± 0.5 attomoles. Quantification of miR-21 from human tissues and plasma samples revealed the relevance for breast and ovarian cancer diagnostics. Analysis of miR-132 and miR-146a from acute monocytic leukemia cells (THP-1) demonstrated the broad applicability to different miRNAs and other types of clinical samples. Direct comparison to the gold standard RT-qPCR showed advantages of amplified TG-FRET concerning precision and specificity when quantifying low concentrations of miRNAs as required for diagnostic applications. Our results demonstrate that a careful implementation of rolling circle amplification and TG-FRET into one straightforward nucleic acid detection method can significantly advance the possibilities of miRNA-based cancer diagnostics and research.

Graphical abstract: Advanced microRNA-based cancer diagnostics using amplified time-gated FRET

Supplementary files

Article information

Article type
Edge Article
14 Jul 2018
10 Sep 2018
First published
11 Sep 2018
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2018,9, 8046-8055

Advanced microRNA-based cancer diagnostics using amplified time-gated FRET

X. Qiu, J. Xu, J. Guo, A. Yahia-Ammar, N. Kapetanakis, I. Duroux-Richard, J. J. Unterluggauer, N. Golob-Schwarzl, C. Regeard, C. Uzan, S. Gouy, M. DuBow, J. Haybaeck, F. Apparailly, P. Busson and N. Hildebrandt, Chem. Sci., 2018, 9, 8046 DOI: 10.1039/C8SC03121E

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