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Single cell immunodetection of Escherichia coli O157:H7 on an indium-tin-oxide electrode by using an electrochemical label with an organic-inorganic nanostructure

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Abstract

A rapid and highly sensitive method is described for the detection of enterohemorrhagic Escherichia coli O157:H7. An organic-inorganic nanostructure in which numerous gold nanoparticles (AuNPs) are enclosed with polyaniline (PANI) was utilized as an electrochemical label. The nanostructure showed (a) strong light scattering intensity due to the coupling effect of the surface plasmon resonance based on the presence of AuNPs, and (b) high electrochemical response due to the redox activity of PANI. To achieve selectivity, antibody against E. coli O157:H7 was immobilized on the surface of the nanostructure. The method exploits the combination of strong adsorption of bacterial cells onto the indium-tin-oxide (ITO) glass electrode without any special processing and specific binding of the nanostructured label to E. coli O157:H7. This enables the electrochemical detection of a single cell on the ITO electrode. The electrochemical response to E. coli O157:H7 was 30-fold higher than that to other types of bacteria. This procedure can be applied to the determination of E. coli O157:H7 even in the presence of other bacteria.

Schematic of a voltammetric immunoassay for Escherichia coli O157:H7 by using a nanocomposite consisting of gold nanoparticles and polyaniline on an ITO electrode

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Acknowledgements

The authors thank Professor M. Miyake at Osaka Prefecture University and Dr. K. Seto at Osaka Prefectural Institute of Public Health for providing verotoxin-non-producing E. coli strains. This work was financially supported by the Ministry of Agriculture, Forestry, and Fisheries through a science and technology research promotion program for the agriculture, forestry, fisheries, and food industries; and the Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research (B) (KAKENHI 25288039, 16H04137) and Grant-in-Aid for Challenging Exploratory Research (KAKENHI 26620072).

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  1. Department of Applied Chemistry, Osaka Prefecture University, 1-2 Gakuen, Naka, Sakai, Osaka, 599-8570, Japan

    Dung Quang Nguyen, Kengo Ishiki & Hiroshi Shiigi

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  1. Dung Quang Nguyen
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Correspondence to Hiroshi Shiigi.

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Nguyen, D.Q., Ishiki, K. & Shiigi, H. Single cell immunodetection of Escherichia coli O157:H7 on an indium-tin-oxide electrode by using an electrochemical label with an organic-inorganic nanostructure. Microchim Acta 185, 465 (2018). https://doi.org/10.1007/s00604-018-3001-5

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