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

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Abstract

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

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Publication details

The article was received on 14 Jul 2018, accepted on 10 Sep 2018 and first published on 11 Sep 2018


Article type: Edge Article
DOI: 10.1039/C8SC03121E
Citation: Chem. Sci., 2018, Advance Article
  • Open access: Creative Commons BY license
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    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, Advance Article , DOI: 10.1039/C8SC03121E

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