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Integrated nicking enzyme-powered numerous-legged DNA walker prepared by rolling circle amplification for fluorescence detection of microRNA

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Abstract

MicroRNAs (miRNAs) have been accepted as promising non-invasive biomarkers for cancer early diagnosis. Developing amplified sensing strategies for detecting ultralow concentration of miRNAs in clinical samples still requires much effort. Herein, an integrated fluorescence biosensor using nicking enzyme-powered numerous-feet DNA walking machine was developed for ultrasensitive detection of miRNA. A long numerous-feet walker produced by target-triggered rolling circle amplification autonomously moves along the defined DNA tracks on gold nanorods (AuNRs) with the help of nicking enzyme, leading to the recovery of fluorescence. This results in an amplified fluorescence signal, typically measured at 518 nm emission wavelength. Benefiting from the long walker that dramatically improves movement range, the homogenous and one-step strategy realizes ultrahigh sensitivity with a limit of detection of 0.8 fM. Furthermore, this walking machine has been successfully used to quantification of miRNA in clinical serum samples. The consistency of the gained results between of the developed strategy and reverse transcription quantitative polymerase chain reaction (RT-qPCR) shows that the sensing method has great promise for tumor diagnostics based on nucleic acid.

Graphical abstract

Schematic representation of the fluorescent biosensing strategy, numerous-legged DNA walker prepared by rolling circle amplification on gold nanorods (AuNRs) for microRNA analysis, which can be applied in real samples with good results.

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Funding

This work was supported by the Natural Science Foundation Project of Chongqing (cstc2020jcyj-msxmX0611) and the Talent Fund of the Fengjie People’s Hospital.

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Author notes

  1. Lihua Wang and Hanqing Zeng contributed equally to this work.

Authors and Affiliations

  1. Health Management Medical Examination Center, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, 404600, China

    Lihua Wang

  2. Department of Hematology, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, 404600, China

    Hanqing Zeng

  3. Department of Neurology, the Fengjie People’s Hospital, Fengjie Branch of the Second Affiliated Hospital of Chongqing Medical University, Fengjie County, Chongqing, 404600, China

    Xiaolan Yang, Chaoming Chen & Shu Ou

Authors
  1. Lihua Wang
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  2. Hanqing Zeng
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  3. Xiaolan Yang
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  4. Chaoming Chen
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  5. Shu Ou
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Correspondence to Shu Ou.

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Wang, L., Zeng, H., Yang, X. et al. Integrated nicking enzyme-powered numerous-legged DNA walker prepared by rolling circle amplification for fluorescence detection of microRNA. Microchim Acta 188, 214 (2021). https://doi.org/10.1007/s00604-021-04875-1

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  • DOI: https://doi.org/10.1007/s00604-021-04875-1

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