Original Paper

Analytical and Bioanalytical Chemistry

, Volume 401, Issue 8, pp 2549-2559

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

  • Stuart W. J. EmberAffiliated withDivision of Pathway Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh
  • , Holger SchulzeAffiliated withDivision of Pathway Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh
  • , Alan J. RossAffiliated withDivision of Pathway Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh
  • , Julie LubyAffiliated withDivision of Pathway Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh
  • , Mizanur KhondokerAffiliated withDivision of Pathway Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh
  • , Gerard GiraudAffiliated withSchool of Physics and Astronomy, The University of Edinburgh
  • , Jonathan G. TerryAffiliated withInstitute of Integrated Systems, School of Engineering, The University of Edinburgh
  • , Ilenia CianiAffiliated withEaStCHEM School of Chemistry, The University of Edinburgh
  • , Chaker TliliAffiliated withEaStCHEM School of Chemistry, The University of Edinburgh
    • , Jason CrainAffiliated withSchool of Physics and Astronomy, The University of EdinburghNational Physical Laboratory
    • , Anthony J. WaltonAffiliated withInstitute of Integrated Systems, School of Engineering, The University of Edinburgh
    • , Andrew R. MountAffiliated withEaStCHEM School of Chemistry, The University of Edinburgh
    • , Peter GhazalAffiliated withDivision of Pathway Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh
    • , Till T. BachmannAffiliated withDivision of Pathway Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh
    • , Colin J. CampbellAffiliated withDivision of Pathway Medicine, College of Medicine and Veterinary Medicine, The University of EdinburghEaStCHEM School of Chemistry, The University of Edinburgh Email author 

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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.

Keywords

DNA microarray Protein microarray HCV Time-to-result Point-of-care Seroconversion panel