A simple and rapid method for the quantitation of secreted hepatitis B virions in cell culture models.

Cell culture models for hepatitis B virus (HBV) remain the mainstay for screening and testing the efficacy of anti‑hepatitis B virus agents. Gradient‑based ultracentrifugation followed by Southern Blotting is used for hepatitis B virion estimation in cell culture; this method has several limitations. We report the development of an assay using a commercially available HBsAg‑ELISA plate for immunocapture followed by real‑time PCR for quantification of hepatitis B virions in cell cultures. This assay is rapid, highly sensitive (50 copies/reaction) and highly specific for virion‑associated DNA. In addition, the assay requires only 20 μL of supernatant, allowing scaling down of transfections.