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Abstract Details
An adaptable platform for in-house hepatitis C serology
1Département de Microbiologie-Infectiologie et Immunologie, Faculté de Médecine, Université Laval, Québec, Canada.
2Centre Hospitalier Universitaire de Libreville, Libreville BP2228, Gabon.
3Department of Infectious Diseases, Copenhagen University Hospital, Hvidovre, Denmark; Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
4Unité d'Evolution Epidémiologie et Résistances Parasitaires, Centre Interdisciplinaire de Recherches Médicales de Franceville, Franceville, Gabon.
5Axe des Maladies Infectieuses et Immunitaires, Centre de Recherche du Centre Hospitalier Universitaire de Québec-Université Laval, Québec G1V 4G2, Canada.
6Galveston National Laboratory, Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA.
7Département de Microbiologie-Infectiologie et Immunologie, Faculté de Médecine, Université Laval, Québec, Canada; Global Urgent and Advanced Research and Development - GUARD, 911 Rue Principale, unit 100, Batiscan, Quebec G0X 1A0, Canada. Electronic address: faustlbv@gmail.com.
Abstract
Serology-based diagnosis remains one of the major tools for diagnosis and surveillance of infectious diseases. However, for many neglected diseases no or only few commercial assays are available and often with prices prohibiting large scale testing in low and middle-income countries (LMICs). We developed an adaptable enzyme-linked immunoassay (ELISA) using hepatitis C virus (HCV) as a proof-of-concept application. By combining the maltose-binding-protein with a multiepitope HCV protein, we were able to obtain a high concentration of protein suitable for downstream applications. Following optimization, the assay was verified using previously tested human samples from Canada, Denmark and Gabon in parallel with the use of a commercial protein. Sensitivity and specificity were calculated to 98 % and 97 % respectively, after accounting for non-specific binding and assay optimization. This study provides a thorough description of the development, and validation of a multiepitope ELISA-based diagnostic assay against HCV, which could be implemented at low cost. The described methodology can be readily adapted to develop novel ELISA-based diagnostic assays for other infectious pathogens with well-described immunogenic epitopes. This method could improve the diagnosis of neglected diseases for which affordable diagnostic assays are lacking.