Journal of Clinical Microbiology, June 1999, p. 1802-1808, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Diagnostic Division,
Received 6 October 1998/Returned for modification 19 November
1998/Accepted 16 March 1999
A highly sensitive enzyme immunoassay (EIA) for the hepatitis C
virus (HCV) core antigen (HCVcAg) was developed, and its performance was compared with that of the AMPLICOR HCV test (Roche Molecular Systems). The developed one-step pretreatment method, 30-min incubation of the specimen with a solution containing three different types of
detergents (Triton X-100,
3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate [CHAPS],
and sodium dodecyl sulfate), does not require any special device.
Because the interfering anti-core antibody in the sample was
sufficiently inactivated by the pretreatment, HCVcAg in the sample
could be detected. The immunoreactivity on gel filtration was shifted
from void fractions to those corresponding to the molecular mass range
from 20 to 25 kDa, which is equal to the estimated molecular mass of
HCVcAg, after the pretreatment. By the recovery test with
HCVcAg-positive serum, the recovery rate was 93.5 to 106.5%. There
was no interference with the EIA by anticoagulants or blood components
in the serum. When the cutoff value was tentatively set at 0.5 mU/ml
based on the distribution of healthy subjects' sera, the sera of all
healthy subjects (n = 125) and patients with hepatitis
B (n = 50) were negative. HCVcAg was detected in
sera from 57 of 73 individuals (78.1%) with anti-HCV antibody.
Similarly, HCV RNA was detected in sera from 59 individuals (80.8%)
with the AMPLICOR HCV as the qualitative test (AMPLICOR HCV test) and
in sera from 54 individuals (74.0%) by the AMPLICOR HCV Monitor as the
quantitative test (AMPLICOR Monitor test). Concentrations of HCVcAg
and HCV RNA (measured by the AMPLICOR Monitor test) correlated
significantly (r = 0.8, P < 0.001).
On seroconversion panels, HCVcAg was detected during the early
stage of infection, when anti-HCV antibodies had not been produced. This assay for HCVcAg is simpler than assays for HCV RNA based on
gene technology and shows specificity and sensitivity equivalent to
those of the AMPLICOR HCV test.
*
Corresponding author. Mailing address: Diagnostic
Division, Tonen Corporation, 1-3-1 Nishi-tsurugaoka, Ohi-Machi,
Iruma-Gun, Saitama 356-8505, Japan. Phone: 81 492-66-8327. Fax: 81 492-66-8385. E-mail: katsumi.aoyagi{at}tonen.co.jp.
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