The VITROS antihepatitis A virus (anti-HAV) total assay is a competitive immunoassay method based on the preincubation of anti-HAV total in the sample with HAV antigen in the assay reagent, followed by incubation with horseradish peroxidase (HRP)-labeled antibody conjugate (mouse monoclonal anti-HAV). The immune complex is captured by streptavidin on the well surface. Unbound conjugate is removed by washing.
The bound HRP conjugate is measured by a luminescent reaction. A reagent containing luminogenic substrates (a luminol derivative and a peracid salt) and an electron transfer agent is added to the wells. The HRP in the bound conjugate catalyzes the oxidation of the luminol derivative, producing light. The electron transfer agent increases the level and duration of the light produced. The light signals are read by the VITROS Immunodiagnostic system. The amount of HRP conjugate is indicative of the concentration of anti-HAV present in the sample.(Package insert: VITROS Anti-HAV Total Reagent Pack, no. GEM1235A, version 4.0, Ortho-Clinical Diagnostics, Inc. Rochester, NY 14626-5101, 1/22/2012)