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

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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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Authors and Affiliations

  1. Department of Clinical Laboratory Medicine, Southwest Hospital, Third Military Medical University (Army Medical University), 30 Gaotanyan, Shapingba District, Chongqing, 400038, China

    Xiaoqi Tang, Yang Wang, Lin Zhou, Wenqing Zhang, Sha Yang, Lianyu Yu, Shuang Zhao, Kai Chang & Ming Chen

  2. College of Pharmacy and Laboratory Medicine, Third Military Medical University (Army Medical University), 30 Gaotanyan, Shapingba District, Chongqing, 400038, China

    Yang Wang & Ming Chen

  3. State Key Laboratory of Trauma, Burn and Combined Injury, Third Military Medical University (Army Medical University), 30 Gaotanyan, Shapingba District, Chongqing, 400038, China

    Ming Chen

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  1. Xiaoqi Tang
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  2. Yang Wang
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  3. Lin Zhou
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  9. Ming Chen
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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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