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Development of a microtitre plate assay and a rapid membrane-based assay for the detection of antibodies to hepatitis C virus (HCV) in human serum

Tymon, Gráinne (2000) Development of a microtitre plate assay and a rapid membrane-based assay for the detection of antibodies to hepatitis C virus (HCV) in human serum. Master of Science thesis, Dublin City University.

Hepatitis C vims (HCV) was discovered in 1989 and was identified as the causative agent of non-A, non-B hepatitis. Third-generation ELISA techniques, incorporating the use of antigens specific to all hepatitis C viral regions, are used to diagnose HCV infection by serum antibody determination. Rapid immunoblot assay (RIBA) testing identifies antibodies specific to structural and non-structural viral proteins. RT-PCR is established as the gold standard of HCV RNA detection. A positive result on the developed assays does not indicate that the hepatitis C virus is present and replicating. A confirmatory test is needed to assess current HCV status and to monitor treatment. The development of qualitative anti-HCV immunoglobulin G antibody tests is described in this study. A microtitre plate assay, suitable for multiple sample screening was optimised for use in standard laboratories. The rapid membrane-based assay, developed for use at point-of-care, requires no sample preparation. It is a user-friendly assay requiring no laboratory equipment or operator training. Both formats utilise bound synthetic peptides specific for non-structural and core regions of the HCV genome. Optimisation of assays was based on testing with characterised sera. Performance evaluation of the plate assay was carried out on 530 samples achieving 97.9% specificity and 93.6% sensitivity. A total of 751 samples evaluated on the rapid assay resulted in 99.4% specificity and 98.9% sensitivity.
Item Type:Thesis (Master of Science)
Date of Award:2000
Supervisor(s):Balbirnie, Edward
Uncontrolled Keywords:Viral antibodies Detection; Hepatitis C virus
Subjects:Biological Sciences > Biotechnology
DCU Faculties and Centres:DCU Faculties and Schools > Faculty of Science and Health > School of Biotechnology
Use License:This item is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 License. View License
ID Code:19447
Deposited On:03 Oct 2013 10:51 by Celine Campbell . Last Modified 03 Oct 2013 10:51

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