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Sandwich-type microRNA biosensor based on graphene oxide incorporated 3D-flower-like MoS2 and AuNPs coupling with HRP enzyme signal amplification

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Abstract

A sandwich electrochemical biosensing strategy for ultrasensitive detection of miRNA-21 was developed by using graphene oxide incorporated 3D-flower-like MoS2 (3D MoS2-rGO) nanocomposites as the substrate and horseradish peroxidase (HRP)-functionalized DNA strand 1 (S1)-gold nanoparticles (S1-AuNPs-HRP) as signal amplification probes. Herein, 3D MoS2-rGO nanocomposites not only had a large specific surface area and excellent conductivity, but also provided more attachment sites for electrodepositing AuNPs. In the presence of target miRNA, a sandwich structure was formed, and the determination of the miRNA-21 was carried out by measuring the DPV response of H2O2 mediated by hydroquinone (HQ) at a potential of + 0.052 V (vs AgCl reference electrode). Under the optimal experimental conditions, the as-prepared biosensor enabled the ultrasensitive detection of miRNA-21 from 5 fM to 0.5 μM with the low detection limit of 0.54 fM (S/N = 3), comparable or lower than previous reported methods for miRNA-21 detection, which benefited from the synergistic amplification of 3D MoS2-rGO and AuNPs-HRP. The prepared biosensor showed satisfactory selectivity, reproducibility, and stability towards miRNA-21 detection. The biosensor was feasible for accurate and quantitative detection of miRNA-21 in normal human serum samples with RSD below 5.86%, which showed a great potential in clinical analysis and disease diagnosis.

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Funding

This work was supported by the National Natural Science Foundation of China (NO. 81772290), Graduate Scientific Research and Innovation Foundation of Chongqing, China (CYS21069, 2020CDCGJ014), Fundamental Research Funds for the Central Universities (2021CDJYGRH006), National Facility for Translational Medicine (Shanghai) Open Research Fund (TMSK-2021–113), Science and Technology Research Program of Chongqing Education Commission of China (KJCXZD2020008), Chongqing Graduate Tutor Team Construction Project, Analytical and Testing Center of Chongqing University for (FE-SEM / XPS / XRD) and the sharing fund of Chongqing University’s large equipment.

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Authors and Affiliations

  1. Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, Bioengineering College of Chongqing University, Chongqing, 400044, People’s Republic of China

    Jiangbo Dong, Huisi Yang, Jiaying Zhao, Li Wen, Congjuan He, Zhikun Hu, Danqun Huo & Changjun Hou

  2. Chongqing University Three Gorges Hospital, Chongqing, 404000, People’s Republic of China

    Jiawei Li

  3. Chongqing Key Laboratory of Bio-Perception & Intelligent Information Processing, School of Microelectronics and Communication Engineering, Chongqing University, Chongqing, 400044, People’s Republic of China

    Jiawei Li & Danqun Huo

  4. National Facility for Translational Medicine (Shanghai), Shanghai, 200240, People’s Republic of China

    Changjun Hou

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  1. Jiangbo Dong
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  2. Huisi Yang
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  3. Jiaying Zhao
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  4. Li Wen
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Correspondence to Jiawei Li, Danqun Huo or Changjun Hou.

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Dong, J., Yang, H., Zhao, J. et al. Sandwich-type microRNA biosensor based on graphene oxide incorporated 3D-flower-like MoS2 and AuNPs coupling with HRP enzyme signal amplification. Microchim Acta 189, 49 (2022). https://doi.org/10.1007/s00604-021-05141-0

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