Enhanced sensitivity of a second generation ELISA for antibody to hepatitis C virus.
A second generation ELISA for combined detection of antibodies to three hepatitis C virus (HCV) recombinant proteins, i.e. C100, C33c and core, was compared with a first generation anti-HCV ELISA in which only antibodies to C100 are detected. The results of the ELISAs were evaluated in 225 haemophilia patients (panel A) and 44 patients with non-A, non-B (NANB) hepatitis (panel B). HCV infection was established by cDNA-polymerase chain reaction (PCR) (in panel B only) and by studying the anti-HCV reaction patterns in 4 separate ELISAs for detection of antibodies to the recombinant proteins C100, C33c, core and a combination of two synthetic peptides sp67/65 derived from the C100 region. The sensitivity for the detection of HCV infection had increased from 0.92[95% confidence interval (CI): 0.87-0.95] to 1.00 (95% CI: 0.89-1.00) in haemophiliacs and from 0.84 (95% CI: 0.66-0.95) to 1.00 (95% CI: 0.89-1.00) in NANB hepatitis patients when the second generation ELISA was used instead of the first generation ELISA. Concurrently the chance of a false negative result was reduced in panel A and B from 0.37 to 0 and from 0.28 to 0, respectively. Analysis of anti-HCV reaction patterns revealed that 172 of 206 (83.5%) anti-HCV ELISA-reactive haemophilia patients had antibodies to all 4 antigens tested. In the NANB hepatitis patients 18 of 31 (58.1%) anti-HCV ELISA-reactive subjects reacted with 4 antigens. In the PCR tested panel of NANB hepatitis patients 2 subjects who showed antibody reactivity to only one antigen and 5 patients with reactivity to 2 antigens were PCR-positive.(ABSTRACT TRUNCATED AT 250 WORDS)
Bresters
D
D
Central Laboratory of The Netherlands, Red Cross Blood Transfusion Service, Amsterdam, The Netherlands.
Cuypers
H T
HT
Reesink
H W
HW
Schaasberg
W P
WP
van der Poel
C L
CL
Mauser-Bunschoten
E P
EP
Houghton
M
M
Choo
Q L
QL
Kuo
G
G
Lesniewski
R
R
D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005188:False Negative Reactions; D006467:Hemophilia A / Q000150:complications / Q000276:immunology / Q000628:therapy; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology*; D006508:Hepatitis Antibodies / Q000032:analysis*; D006526:Hepatitis C / Q000097:blood / Q000453:epidemiology / Q000209:etiology / Q000276:immunology* / Q000635:transmission; D018936:Hepatitis C Antigens; D006801:Humans; D008297:Male; D010446:Peptide Fragments / Q000276:immunology; D016133:Polymerase Chain Reaction; D015995:Prevalence; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D065227:Transfusion Reaction; D014758:Viral Core Proteins / Q000276:immunology; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology
D000956:Antigens, Viral; C064544:C100 protein, hepatitis C virus; D006508:Hepatitis Antibodies; D018936:Hepatitis C Antigens; D010446:Peptide Fragments; D011994:Recombinant Proteins; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; C069130:nucleocapsid protein, Hepatitis C virus