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Integration of isothermal amplification with quantum dot-based fluorescence resonance energy transfer for simultaneous detection of multiple microRNAs

Abstract

MicroRNAs (miRNAs) are small non-coding RNAs that regulate important physiological processes, and their dysregulation is associated with various human diseases. Simultaneously sensitive detection of multiple miRNAs may facilitate the early clinical diagnosis. In this research, we demonstrate for the first time the integration of hyperbranched rolling circle amplification (HRCA) with quantum dot (QD)-based fluorescence resonance energy transfer (FRET) for simultaneous detection of multiple microRNAs with single-color QD as the donor and two fluorescent dyes as the acceptors. We used miR-21 and miR-221 as the target miRNAs. We designed two circular templates which may specifically hybridize with miR-21 and miR-221, respectively, for the initiation of HRCA reaction. The products of HRCA reaction may hybridize with both capture probes and reporter probes to form the biotinylated acceptor-labeled sandwiched hybrids. The resultant sandwiched hybrids can assemble on the surface of QD, enabling efficient FRET between the QD and the acceptors, with the Cy3 signal indicating the presence of miR-21 and the Texas Red signal indicating the presence of miR-221. This assay has significant advantages of simplicity and low cost. The HRCA reaction can be performed under isothermal condition with same reverse primer for different target miRNAs, and the products of HRCA reaction for both miR-21 and miR-221 can specifically hybridize with the same capture probes. This assay exhibits excellent specificity and high sensitivity with a detection limit of 7.2 × 10−16 M for miR-21 and 1.6 × 10−17 M for miR-221, and it can be used for simultaneous detection of multiple miRNAs in human cancer cells, holding great potential in biomedical research and clinical diagnosis.

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Publication details

The article was received on 20 Feb 2018, accepted on 10 Apr 2018 and first published on 12 Apr 2018


Article type: Edge Article
DOI: 10.1039/C8SC00832A
Citation: Chem. Sci., 2018, Accepted Manuscript
  • Open access: Creative Commons BY license
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    Integration of isothermal amplification with quantum dot-based fluorescence resonance energy transfer for simultaneous detection of multiple microRNAs

    J. Hu, M. Liu and C. Zhang, Chem. Sci., 2018, Accepted Manuscript , DOI: 10.1039/C8SC00832A

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