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Updated: Aug 20, 2026
A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
Novel assay using total hepatitis C virus (HCV) core antigen quantification for diagnosis of HCV infection in
Fabrizio Fabrizi1, Giovanna Lunghi, Filippo Aucella
1Division of Nephrology, Maggiore Hospital, IRCCS, via Commenda 15, 20122 Milan, Italy. fabrizi@policlinico.mi.it
Insights
A new assay measuring total hepatitis C virus (HCV) core antigen (Ag) correlates well with HCV RNA levels in dialysis patients. This simpler method can aid viral load assessment in routine labs.
Area of Science:
- Hepatology
- Virology
- Clinical Diagnostics
Background:
- Dialysis patients are at high risk for hepatitis C virus (HCV) infection.
- Current HCV diagnosis uses serology and nucleic acid amplification technology (NAT), but NAT is costly and complex.
- A novel assay to detect and quantify total HCV core antigen (Ag) has been developed.
Purpose of the Study:
- To compare a total HCV core Ag enzyme-linked immunosorbent assay (ELISA) with a quantitative HCV RNA assay.
- To evaluate the utility of total HCV core Ag as a surrogate marker for HCV viral load in dialysis patients.
Main Methods:
- A cohort of 305 dialysis patients, positive and negative for HCV, was studied.
- Total HCV core Ag was measured using ELISA.
- HCV RNA levels were quantified using reverse transcription-PCR (RT-PCR).
Main Results:
- Concentrations of total HCV core Ag and HCV RNA levels showed significant correlation (r = 0.471, P = 0.0001).
- This correlation was maintained across different HCV genotypes (1 and 2).
- An estimated equivalence of 1 pg/mL total HCV core Ag to approximately 19,952 IU/mL HCV RNA was determined.
Conclusions:
- Total HCV core Ag measurement is a useful tool for assessing HCV viral load in dialysis patients.
- The total HCV core Ag ELISA can be employed in routine laboratories without specialized equipment or training.
- This supports the use of total anti-HCV core Ag ELISA for viral load assessment in HCV-infected dialysis patients.
Abstract:
Dialysis patients remain a high-risk group for hepatitis C virus (HCV) infection. The current diagnosis of HCV infection among dialysis patients includes serological assays and nucleic acid amplification technology (NAT) for assessing serum anti-HCV antibody and HCV viremia, respectively. However, current NAT techniques are expensive and labor-intensive and often lack standardization. An assay prototype designed to detect and quantify total HCV core antigen (total HCV core Ag) protein in serum and plasma in the presence or absence of anti-HCV antibodies has been recently developed. A comparison between a total anti-HCV core Ag enzyme-linked immunosorbent assay (ELISA) and a quantitative HCV RNA assay based on reverse transcription-PCR (RT-PCR) (Amplicor HCV Monitor test) was performed using a large (n = 305) cohort of ELISA HCV 3.0 HCV-negative and -positive patients on maintenance dialysis. The concentrations of HCV core Ag and HCV RNA levels (measured by RT-PCR) were significantly correlated (r = 0.471, P = 0.0001) over a wide range of HCV RNA levels and were maintained among different HCV genotypes (HCV genotype 1, r = 0.862, P = 0.0001; HCV genotype 2, r = 0.691, P = 0.0001). We estimated that 1 pg of total HCV core Ag per ml is equivalent to approximately 19.952 IU of HCV RNA per ml, even if the wide range in the ratio of core Ag to HCV RNA (95% confidence intervals, 2.8 x 10(3) to 1.6 x 10(5) IU/ml) precluded definitive conclusions. In summary, total HCV core Ag proved to be useful for performing HCV RNA measurement among dialysis patients in routine laboratories without the need for special equipment or training. The present study supports the use of the total anti-HCV core Ag ELISA for assessing viral load among dialysis patients with HCV infection.
