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New-generation RIBA hepatitis C strip immunoblot assays
A J Polito1, R K DiNello, S Quan
1Chiron Corporation, Emeryville, Calif.
Beitrage Zur Infusionstherapie = Contributions to Infusion Therapy
|January 1, 1992
Summary
Second-generation hepatitis C virus (HCV) ELISAs and RIBA assays show improved detection but increased indeterminate results. Further testing is crucial for accurate anti-HCV antibody diagnosis in blood screening.
Area of Science:
- Virology
- Immunology
- Diagnostic Assays
Background:
- Second-generation hepatitis C virus (HCV) ELISAs incorporate additional antigens (nucleocapsid and NS-3 regions) beyond first-generation assays (c100-3 antigen).
- A supplementary second-generation RIBA HCV strip immunoblot assay (2-RIBA HCV SIA) utilizes four recombinant HCV antigens: 5-1-1 (NS-4), c100-3 (NS-4), c33c (NS-3), and c22-3 (NS-3/nucleocapsid).
Purpose of the Study:
- To evaluate the performance of second-generation HCV diagnostic assays, specifically the Ortho second-generation HCV ELISA (ORTHO HCV 2.0 ELISA) and the 2-RIBA HCV SIA.
- To analyze changes in the repeat reactive donor population and the characteristics of indeterminate results generated by these new assays.
Main Methods:
- Screening of random volunteer blood donors using the ORTHO HCV 2.0 ELISA.
- Confirmation and characterization of reactive samples using the 2-RIBA HCV SIA with multiple HCV antigens.
- Analysis of indeterminate results from the 2-RIBA HCV SIA, comparing them to first-generation assay results.
Main Results:
- The ORTHO HCV 2.0 ELISA detected a significant number of samples reactive with second-generation antigens (c33c, c22-3) in the 2-RIBA HCV SIA.
- A higher percentage of ORTHO HCV 2.0 ELISA reactive specimens were found indeterminate by the 2-RIBA HCV SIA (approx. 25%) compared to first-generation assays (approx. 5%).
- Indeterminate samples in second-generation assays showed a shift in reactivity dominance from c100-3 to c22-3.
Conclusions:
- Second-generation HCV assays offer enhanced detection capabilities but yield a higher rate of indeterminate results, necessitating further investigation for accurate diagnosis.
- Resolution of indeterminate samples is critical for both clinical diagnostics and blood donor screening, particularly when donor notification is required.
- Early studies with HCV peptides suggest that nucleocapsid and NS-4 regions provide specific anti-HCV antibody evidence, while NS-3 (c33c) epitopes may be conformational and difficult to detect with current peptide-based methods.