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Label-free ITO-based immunosensor for the detection of very low concentrations of pathogenic bacteria.
Bioelectrochemistry. 2015 Feb; 101:146-52.B

Abstract

Here we describe the fabrication of a highly sensitive and label-free ITO-based impedimetric immunosensor for the detection of pathogenic bacteria Escherichia coli O157:H7. Anti-E. coli antibodies were immobilized onto ITO electrodes using a simple, robust and direct methodology. First, the covalent attachment of epoxysilane on the ITO surface was demonstrated by Atomic Force Microscopy and cyclic voltammetry. The immobilization of antibody on the epoxysilane layer was quantified by Optical Waveguide Lightmode Spectroscopy, obtaining a mass variation of 12 ng cm(− 2) (0.08 pmol cm(− 2)). Microcontact printing and fluorescence microscopy were used to demonstrate the specific binding of E. coli O157:H7 to the antibody-patterned surface. We achieved a ratio of 1:500 Salmonella typhimurium/E. coli O157:H7, thus confirming the selectivity of the antibodies and efficiency of the functionalization procedure. Finally, the detection capacity of the ITO-based immunosensor was evaluated by Electrochemical Impedance Spectroscopy. A very low limit of detection was obtained (1 CFU mL(− 1)) over a large linear working range (10–10(6) CFU mL(− 1)). The specificity of the impedimetric immunosensor was also examined. Less than 20% of non-specific bacteria (S. typhimurium and E. coli K12) was observed. Our results reveal the applicability of ITO for the development of highly sensitive and selective impedimetric immunosensors.

Authors

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Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25460610

Citation

Barreiros dos Santos, M, et al. "Label-free ITO-based Immunosensor for the Detection of Very Low Concentrations of Pathogenic Bacteria." Bioelectrochemistry (Amsterdam, Netherlands), vol. 101, 2015, pp. 146-52.
Barreiros dos Santos M, Azevedo S, Agusil JP, et al. Label-free ITO-based immunosensor for the detection of very low concentrations of pathogenic bacteria. Bioelectrochemistry. 2015;101:146-52.
Barreiros dos Santos, M., Azevedo, S., Agusil, J. P., Prieto-Simón, B., Sporer, C., Torrents, E., Juárez, A., Teixeira, V., & Samitier, J. (2015). Label-free ITO-based immunosensor for the detection of very low concentrations of pathogenic bacteria. Bioelectrochemistry (Amsterdam, Netherlands), 101, 146-52.
Barreiros dos Santos M, et al. Label-free ITO-based Immunosensor for the Detection of Very Low Concentrations of Pathogenic Bacteria. Bioelectrochemistry. 2015;101:146-52. PubMed PMID: 25460610.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Label-free ITO-based immunosensor for the detection of very low concentrations of pathogenic bacteria. AU - Barreiros dos Santos,M, AU - Azevedo,S, AU - Agusil,J P, AU - Prieto-Simón,B, AU - Sporer,C, AU - Torrents,E, AU - Juárez,A, AU - Teixeira,V, AU - Samitier,J, PY - 2014/05/20/received PY - 2014/09/20/revised PY - 2014/09/21/accepted PY - 2014/12/3/entrez PY - 2014/12/3/pubmed PY - 2016/4/29/medline SP - 146 EP - 52 JF - Bioelectrochemistry (Amsterdam, Netherlands) JO - Bioelectrochemistry VL - 101 N2 - Here we describe the fabrication of a highly sensitive and label-free ITO-based impedimetric immunosensor for the detection of pathogenic bacteria Escherichia coli O157:H7. Anti-E. coli antibodies were immobilized onto ITO electrodes using a simple, robust and direct methodology. First, the covalent attachment of epoxysilane on the ITO surface was demonstrated by Atomic Force Microscopy and cyclic voltammetry. The immobilization of antibody on the epoxysilane layer was quantified by Optical Waveguide Lightmode Spectroscopy, obtaining a mass variation of 12 ng cm(− 2) (0.08 pmol cm(− 2)). Microcontact printing and fluorescence microscopy were used to demonstrate the specific binding of E. coli O157:H7 to the antibody-patterned surface. We achieved a ratio of 1:500 Salmonella typhimurium/E. coli O157:H7, thus confirming the selectivity of the antibodies and efficiency of the functionalization procedure. Finally, the detection capacity of the ITO-based immunosensor was evaluated by Electrochemical Impedance Spectroscopy. A very low limit of detection was obtained (1 CFU mL(− 1)) over a large linear working range (10–10(6) CFU mL(− 1)). The specificity of the impedimetric immunosensor was also examined. Less than 20% of non-specific bacteria (S. typhimurium and E. coli K12) was observed. Our results reveal the applicability of ITO for the development of highly sensitive and selective impedimetric immunosensors. SN - 1878-562X UR - https://www.unboundmedicine.com/medline/citation/25460610/Label_free_ITO_based_immunosensor_for_the_detection_of_very_low_concentrations_of_pathogenic_bacteria_ DB - PRIME DP - Unbound Medicine ER -