Improved diagnosis of chronic hepatitis C virus infection by detection of antibody to multiple epitopes: confirmation by antibody to synthetic oligopeptides.
Serum samples from 226 patients covering a wide spectrum of liver disease were tested for antibodies to hepatitis C virus (HCV) using both first and second generation enzyme linked immunosorbent assays. Selected sera were also tested by peptide immunoassays, by the four-antigen recombinant immunoblot assay (RIBA II), and for viral genome by the polymerase chain reaction. Antibody to c100-3 was detected in 61% of patients with chronic non-A, non-B (NANB) hepatitis and/or 46.5% with presumed NANB-related cirrhosis by the first generation test. These figures increased to 77% and 58% when antibodies to recombinant structural and non-structural HCV antigens were sought by the second generation assay. Supplemental testing against peptide Sp75 and Sp65/sp67 confirmed that reactivity of sera by second generation assays was due to antibodies to the additional structural and non-structural antigens. Samples negative by the first generation assay were not confirmed by the supplemental assay using peptides Sp75 and Sp65/Sp67. HCV RNA was detected in 60% of the anti-HCV positive sera tested, most of which were also RIBA II positive. Our findings confirm that the introduction of the structural and non-structural antigens, especially the putative nucleocapsid protein, improves sensitivity of detection of antibodies to HCV, and facilitates diagnosis in patients with "cryptogenic" chronic hepatitis.
Brown
D
D
University Department of Medicine, Royal Free Hospital, London, England.
Powell
L
L
Morris
A
A
Rassam
S
S
Sherlock
S
S
McIntyre
N
N
Zuckerman
A J
AJ
Dusheiko
G M
GM
D000956:Antigens, Viral / Q000276:immunology*; D001483:Base Sequence; D002908:Chronic Disease; D004277:DNA, Single-Stranded / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology*; D006508:Hepatitis Antibodies / Q000097:blood*; D006526:Hepatitis C / Q000175:diagnosis*; D018937:Hepatitis C Antibodies; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000276:immunology; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000097:blood; D011994:Recombinant Proteins / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000276:immunology
D000956:Antigens, Viral; C064544:C100 protein, hepatitis C virus; D004277:DNA, Single-Stranded; D000939:Epitopes; D006508:Hepatitis Antibodies; D018937:Hepatitis C Antibodies; D009842:Oligopeptides; D012367:RNA, Viral; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins