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Strand displacement-triggered G-quadruplex/rolling circle amplification strategy for the ultra-sensitive electrochemical sensing of exosomal microRNAs

Abstract

Emerging evidence suggests that exosomal microRNAs are potential biomarkers for the early diagnosis and prognostic assessment of tumor. Here, we design a strand displacement-initiated G-quadruplex/rolling circle amplification (RCA) strategy for highly specific and sensitive electrochemical sensing of exosomal microRNAs. In the presence of exosomal miRNA-21, a locked nucleic acid (LNA)-labeled toehold mediated strand displacement reaction (TMSDR) is initiated, releasing output P2 to trigger the subsequent RCA reaction by hybridizing with the C-rich circular template. Then the obtained G-rich RCA products can bind to the probe anchored on the surface of gold electrode and generate G-quadruplex conformations. Based on the TMSDR-triggered G-quadruplex/RCA strategy, the detection limit of this electrochemical biosensor is down to 2.75 fM. Moreover, our biosensor exhibits excellent repeatability, stability, and high consistency compared to RT-PCR for clinical detection. In conclusion, this assay is expected to provide a hopeful strategy for the early non-invasive diagnosis and prognostic estimation of cancer.

Schematic illustration of electrochemical sensing of exosomal microRNAs based on strand displacement-initiated G-quadruplex/rolling circle amplification (RCA) strategy.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant No. 81430053, 81972027), Chongqing Health Commission (2018QNXM049, 2019ZDXM025) and Medical pre-research project of the Army Medical University (2018XYY04).

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Correspondence to Kai Chang or Ming Chen.

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Tang, X., Wang, Y., Zhou, L. et al. Strand displacement-triggered G-quadruplex/rolling circle amplification strategy for the ultra-sensitive electrochemical sensing of exosomal microRNAs. Microchim Acta 187, 172 (2020). https://doi.org/10.1007/s00604-020-4143-9

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Keywords

  • Exosomal microRNAs
  • Locked nucleic acid
  • Strand displacement
  • Rolling circle amplification
  • G-quadruplex
  • Biosensor