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Fast DNA and protein microarray tests for the diagnosis of hepatitis C virus infection on a single platform

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Abstract

Hepatitis C virus (HCV) is a major cause of chronic liver disease and liver cancer, and remains a large health care burden to the world. In this study we developed a DNA microarray test to detect HCV RNA and a protein microarray to detect human anti-HCV antibodies on a single platform. A main focus of this study was to evaluate possibilities to reduce the assay time, as a short time-to-result (TTR) is a prerequisite for a point-of-care test. Significantly reducing hybridisation and washing times did not impair the assay performance. This was confirmed first using artificial targets and subsequently using clinical samples from an HCV seroconversion panel derived from a HCV-infected patient. We were able to reduce the time required for the detection of human anti-HCV antibodies to only 14 min, achieving nanomolar sensitivity. The protein microarray exhibited an analytical sensitivity comparable to that of commercial systems. Similar results were obtained with the DNA microarray using a universal probe which covered all different HCV genotypes. It was possible to reduce the assay time after PCR from 150 min to 16 min without any loss of sensitivity. Taken together, these results constitute a significant step forward in the design of rapid, microarray-based diagnostics for human infectious disease, and show that the protein microarray is currently the most favourable candidate to fill this role.

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Acknowledgements

This work was supported by ITI Techmedia Scotland/Scottish Enterprise (Glasgow, UK) as part of the BioSensing Platform programme. The authors also wish to thank Kate Templeton and Alison Hardie (Royal Infirmary of Edinburgh, Edinburgh, UK) for the extraction of HCV RNA from clinical samples and seroconversion panel samples. CJC was funded by an EaStCHEM Research Fellowship.

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Authors and Affiliations

  1. Division of Pathway Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh, Chancellor’s Building, Little France Crescent, Edinburgh, EH16 4SB, UK

    Stuart W. J. Ember, Holger Schulze, Alan J. Ross, Julie Luby, Mizanur Khondoker, Peter Ghazal, Till T. Bachmann & Colin J. Campbell

  2. School of Physics and Astronomy, The University of Edinburgh, The King’s Buildings, West Mains Road, Edinburgh, EH9 3JZ, UK

    Gerard Giraud & Jason Crain

  3. National Physical Laboratory, Hampton Road, Teddington, Middlesex, TW11 0LW, UK

    Jason Crain

  4. Institute of Integrated Systems, School of Engineering, The University of Edinburgh, The King’s Buildings, West Mains Road, Edinburgh, EH9 3JF, UK

    Jonathan G. Terry & Anthony J. Walton

  5. EaStCHEM School of Chemistry, The University of Edinburgh, Joseph Black Building, West Mains Road, Edinburgh, EH9 3JJ, UK

    Ilenia Ciani, Chaker Tlili, Andrew R. Mount & Colin J. Campbell

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  1. Stuart W. J. Ember
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Corresponding author

Correspondence to Colin J. Campbell.

Additional information

Stuart W. J. Ember and Holger Schulze contributed equally to the work.

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Ember, S.W.J., Schulze, H., Ross, A.J. et al. Fast DNA and protein microarray tests for the diagnosis of hepatitis C virus infection on a single platform. Anal Bioanal Chem 401, 2549–2559 (2011). https://doi.org/10.1007/s00216-011-5332-5

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  • DOI: https://doi.org/10.1007/s00216-011-5332-5

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