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Homogeneous photoelectrochemical biosensor for microRNA based on target-responsive hydrogel coupled with exonuclease III and nicking endonuclease Nb.BbvCI assistant cascaded amplification strategy

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Abstract

MicroRNAs can serve as biomarkers for many cancers, so it is significant to develop simple and sensitive strategies for microRNAs detection. Photoelectrochemical (PEC) detection has the advantages of simple equipment and high sensitivity. But in conventional PEC DNA sensors, tedious immobilization procedures of photoactive materials and capture probes on electrode surfaces are inevitable. To overcome those limitations, a homogeneous PEC biosensor based on target-responsive hydrogels has been developed (miRNA-155 has been chosen as a model target). PEC signal molecules (TiO2 nanoparticles, TiO2 NPs) were embedded in DNA hydrogels formed by hyaluronic acid sodium salt, amine-modified DNA double strands, and polyethylenimine rich in amine groups. In the presence of the target, DNA double strands in hydrogel were nicked by endonuclease and TiO2 NPs were released to the supernate and a high PEC response was obtained when collecting the supernate for PEC test, while almost no TiO2 NPs released in the absence of the target. Thanks to the exonuclease III and nicking endonuclease Nb.BbvCI-assisted cascaded amplification strategy, the proposed biosensor exhibits high sensitivity toward miRNA-155 with a low detection limit of 0.41 fM and a wide linear range from 1.0 fM to 100 pM. Since this method circumvents tedious electrode modification procedures, the proposed technique exhibits the advantages of simplicity and good reproducibility. Moreover, the prepared hydrogels have outstanding storage stability, so that they can be prepared in advance and shorten detection time. This biosensing platform provides a versatile strategy for the construction of homogeneous PEC biosensors for the detection of diverse targets.

Graphical abstract

Photoelectrochemical detection techniques have been coupled with controlled release system to develop an immobilization-free microRNA biosensor. High sensitivity has been realized based on cascaded signal amplification strategy, and the proposed biosensor has been applied to detect the target in real sample with satisfied results. Since no tedious electrode modifications, the proposed homogeneous PEC sensor exhibits high reproducibility and good stability.

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Funding

This project was financially supported by the National Key Research and Development Program of China (No. 2019YFC1604701), National Sciences Foundation of China (21974020, 21775026) and the cooperative project of production and study in University of Fujian Province (2018Y4007), the Sciences Foundation of Fujian Province (2018 J01685, 2018J01682), and the United Fujian Provincial Health and Education Project for Tackling the Key Research P. R. China (2019-WJ-11).

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Authors and Affiliations

  1. Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, 2 Xue Yuan Road, University Town, Fuzhou, Fujian, 350116, People’s Republic of China

    Jiao Yang, Fang Luo, Bin Qiu & Zhenyu Lin

  2. College of Chemistry and Materials Science, Fujian Normal University, Fuzhou, 350117, Fujian, China

    Shilan Fu

  3. College of Biological, Chemical Sciences and Engineering, Jiaxing University, No.56, South Yuexiu Road, Jiaxing, 314001, Zhejiang, People’s Republic of China

    Longhua Guo

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  1. Jiao Yang
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  2. Shilan Fu
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  3. Fang Luo
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  4. Longhua Guo
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  5. Bin Qiu
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Correspondence to Longhua Guo or Zhenyu Lin.

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Yang, J., Fu, S., Luo, F. et al. Homogeneous photoelectrochemical biosensor for microRNA based on target-responsive hydrogel coupled with exonuclease III and nicking endonuclease Nb.BbvCI assistant cascaded amplification strategy. Microchim Acta 188, 267 (2021). https://doi.org/10.1007/s00604-021-04935-6

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