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Voltammetric determination of the Escherichia coli DNA using a screen-printed carbon electrode modified with polyaniline and gold nanoparticles

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Abstract

The authors describe an electrochemical assay for fast detection of Escherichia coli (E. coli). It is based on a dual signal amplification strategy and the use of a screen-printed carbon electrode (SPCE) whose surface was modified with a polyaniline (PANI) film and gold nanoparticles (AuNPs) via cyclic voltammetry (CV). In the next step, avidin was covalently immobilized on the PANI/AuNP composite on the SPCE surface. Subsequently, the biotinylated DNA capture probe was immobilized onto the PANI/AuNP/avidin-modified SPCE by biotin-avidin interaction. Then, DNA of E.coli, digoxigenin-labeled DNA detector probe and anti-digoxigenin-labeled horseradish peroxidase (HRP) were placed on the electrode. 3,3′,5,5′-Tetramethylbenzidine (TMB) and H2O2 solution were added and the CV electrochemical signal was generated at a potential of −0.1 V (vs. Ag/AgCl) and a scan rate 50 mV.s−1. The assay can detect 4 × 106 to 4 CFU of E. coli without DNA amplification. The biosensor is highly specific over other pathogens including Klebsiella pneumoniae, Proteus mirabilis, Enterococcus faecalis, Staphylococcus haemolyticus and Pseudomonas aeruginosa. It can be concluded that this genosensor has an excellent potential for rapid and accurate diagnosis of E.coli inflicted infections.

Schematic of an electrochemical E. coli genosensor based on sandwich assay on a polyaniline/gold nanoparticle-modified screen printed carbon electrode (SPCE). The biosensor can detect 4 × 106 to 4 CFU of E. coli without DNA amplification.

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Acknowledgements

This research was supported by a grant from Tarbiat Modares University.

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Authors and Affiliations

  1. Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, P.O. Box 14115-111, Tehran, Iran

    Nahid Shoaie & Mehdi Forouzandeh

  2. Biosensor Research Center, Endocrinology and Metabolism Molecular-Cellular Sciences Institute, Tehran University of Medical Science, P.O. Box 1411713137, Tehran, Iran

    Mehdi Forouzandeh & Kobra Omidfar

  3. Endocrinology and Metabolism Research Center, Endocrinology and Metabolism Research Institute, Tehran University of Medical Sciences, Tehran, Iran

    Kobra Omidfar

Authors
  1. Nahid Shoaie
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  2. Mehdi Forouzandeh
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  3. Kobra Omidfar
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Correspondence to Mehdi Forouzandeh.

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Shoaie, N., Forouzandeh, M. & Omidfar, K. Voltammetric determination of the Escherichia coli DNA using a screen-printed carbon electrode modified with polyaniline and gold nanoparticles. Microchim Acta 185, 217 (2018). https://doi.org/10.1007/s00604-018-2749-y

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  • DOI: https://doi.org/10.1007/s00604-018-2749-y

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