Abstract
An electrochemical method for the detection of Epstein–Barr virus (EBV) infections is described. The method relies on an immunoassay with electrochemical read-outs based on recombinant antigens. The antigens are immobilised on an Au electrode surface and used to complementarily bind antibodies from serum samples found during different stages of infection with EBV. Thiol chemistry under formation of self-assembled monolayers functions as a means to immobilise the antigens at the Au electrodes. A reporter system consisting of a secondary antibody labelled with alkaline phosphatase is used for electrochemical detection. The feasibility of the assay design is demonstrated and the assay performance is tested against the current gold standard in EBV detection. Close correlation is obtained for the results found for the developed electrochemical immunoassay and a standard line assay. Moreover, the electrochemical immunoassay is combined with a nanoporous electrode system allowing signal amplification by means of redox recycling. An amplification factor of 24 could be achieved.
Amplified immunoassay for the determination of anti-EBV antibodies in serum based on enzyme amplification coupled with electrochemical amplification by redox cycling in nanopore electrodes
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Acknowledgements
The authors are grateful to the German Ministry of Research and Education (BMBF) for funding within the framework of the project NanoBioPore (FKZ 13N8607). Dr. Martin Stelzle, Dr. Ulrich Müller, Prof. Dr. Joachim P. Spatz, Theobald Lohmüller, Dr. Stephan Linke, and Dr. Markus Kaczor are acknowledged for fabrication of nanopore electrode structures and helpful discussions.
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This work was published in the special issue on Focus on Bioanalysis with Guest Editors Antje J. Baeumner, Günter Gauglitz and Frieder W. Scheller.
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Bandilla, M., Zimdars, A., Neugebauer, S. et al. A microelectrochemical sensing system for the determination of Epstein–Barr virus antibodies. Anal Bioanal Chem 398, 2617–2623 (2010). https://doi.org/10.1007/s00216-010-3926-y
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DOI: https://doi.org/10.1007/s00216-010-3926-y