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Proximity-based electrochemical biosensor for highly sensitive determination of methyltransferase activity using gold nanoparticle-based cooperative signal amplification

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Abstract

We describe a turn-on electrochemical biosensor for the detection of methyltransferases (MTases) causing DNA adenine methylation. This biosensor is based on insertion, methylation-resistant cleavage, signal enrichment caused by gold nanoparticles (AuNPs), and a signal probe-dragging strategy. A double-stranded DNA (dsDNA) containing identical MTase and methylation-resistant endonuclease (Mbo I) sites was immobilized on the surface of a gold electrode via Au-S covalent binding. The surface was subsequently treated with MTase and Mbo I and then washed. Results revealed that the surface of the electrode contains methylated dsDNA and 12-base nucleotides residual. Depending on biotin-streptavidin interactions that enabled signal probes and nucleotide residue hybridization and AuNP enrichment, a large number of signal probes labeled with ferrocene (Fc) are captured by the electrode. Under optimal conditions, the differential pulse voltammetry signals of Fc tags (at a working voltage of 0.24 V vs. Ag/AgCl) are linearly related to the log of the MTase activity in the 0.1 to 40 U·mL−1 range. The dynamic range extends from 0.05 to 50 U·mL−1, and the limit of detection is 0.024 U·mL−1 (at an S/N ratio of 3). The assay is well reproducible and highly selective. In our perception, this strategy provides a promising approach for simple, sensitive and selective detection of Dam MTase and may be extended to the determination of other MTase by exchanging the corresponding DNA.

Proximity-based electrochemical biosensor for highly sensitive detection of DNA adenine methylation methyltransferase (Dam MTase) activity using methylation-resistant cleavage coupled with gold nanoparticle based cooperative signal amplification.

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Acknowledgments

This work was financially supported by the Natural Science Research Foundation of China (81171415) and the science & technology bureau of Yibin (2014SF015).

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Authors and Affiliations

  1. Key Laboratory of Laboratory Medical Diagnostics of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, People’s Republic of China

    Zhang Zhang, Xianqing Cao, Juan Yao, Ting Wang & Guoming Xie

  2. The No.2 People’s Hospital of Yibin, Sichuan, 644000, People’s Republic of China

    Shangchun Sheng

  3. Department of Electrical and Computer Engineering, College of Engineering, University of Nevada, Las Vegas, NV, 89154-4026, USA

    Yiyan Li

Authors
  1. Zhang Zhang
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  2. Shangchun Sheng
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  3. Xianqing Cao
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  4. Yiyan Li
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  5. Juan Yao
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  6. Ting Wang
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  7. Guoming Xie
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Correspondence to Guoming Xie.

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Zhang, Z., Sheng, S., Cao, X. et al. Proximity-based electrochemical biosensor for highly sensitive determination of methyltransferase activity using gold nanoparticle-based cooperative signal amplification. Microchim Acta 182, 2329–2336 (2015). https://doi.org/10.1007/s00604-015-1564-y

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  • DOI: https://doi.org/10.1007/s00604-015-1564-y

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