Application of the trak-C HCV core assay for monitoring antiviral activity in HCV replication systems

J Virol Methods. 2004 Jun 1;118(1):23-31. doi: 10.1016/j.jviromet.2004.01.014.

Abstract

The Ortho trak-C immunoassay has recently established detection of the HCV core antigen as a viable indirect marker of HCV replication in clinical samples. In this study, trak-C is used to monitor HCV replication in three pre-clinical models: the cellular HCV replicon system, transient transfection of HCV genomes, and the murine Alb-uPa/SCID HCV infection model. All of these systems utilize full-length HCV genomes that direct the expression of core, facilitating its detection with monoclonal antibodies. When performed with purified protein, the assay detects HCV core with a lower limit of detection at 1.5pg, and exhibits linear detection up to 100pg. When assaying extracts prepared from Huh-7 clone 21-5 cells harboring a full-length HCV replicon, core is detectable from as few as 63 cell equivalents. The assay was used to determine the sensitivity of Huh 21-5 cells to the antiviral effects of interferon (IFN). Inhibition by IFN-alpha using core detection was comparable to that observed using branched-DNA (bDNA 3.0) detection of HCV RNA. Replication of transfected full-length HCV 1a Con1 genomes in Huh-7 cells was also detectable using the trak-C assay. Finally, in the transgenic murine HCV infection model, the course of viral amplification was detected from serum using trak-C with kinetics similar to those observed with RNA detection. Given its ease of use and the lack of requirement for RNA purification, the trak-C assay has several advantages over RNA-based methods of viral monitoring.

MeSH terms

  • Animals
  • Antiviral Agents / pharmacology*
  • Clone Cells
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Enzyme-Linked Immunosorbent Assay / statistics & numerical data
  • Gene Expression / drug effects
  • Genome, Viral
  • Hepacivirus / drug effects*
  • Hepacivirus / genetics
  • Hepacivirus / physiology*
  • Hepatitis C / drug therapy
  • Hepatitis C / virology
  • Humans
  • Interferon Type I / pharmacology
  • Mice
  • Mice, SCID
  • Mice, Transgenic
  • Recombinant Proteins
  • Sensitivity and Specificity
  • Viral Core Proteins / analysis*
  • Viral Core Proteins / genetics
  • Viremia / drug therapy
  • Viremia / virology
  • Virology / methods
  • Virology / statistics & numerical data
  • Virus Replication / drug effects*

Substances

  • Antiviral Agents
  • Interferon Type I
  • Recombinant Proteins
  • Viral Core Proteins
  • nucleocapsid protein, Hepatitis C virus