title: A simple and rapid method for the quantitation of secreted hepatitis B virions in cell culture models.
abstract: 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.
authors: J
authors: J
authors: Samal
authors: M
authors: M
authors: Kandpal
authors: Kusuma School of Biological Sciences, Indian Institute of Technology Delhi, New Delhi, India.
authors: P
authors: P
authors: Vivekanandan
mesh_terms: D004279:DNA, Viral / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006514:Hepatitis B Surface Antigens / Q000276:immunology; D006515:Hepatitis B virus / Q000254:growth & development / Q000302:isolation & purification*; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D019562:Viral Load / Q000379:methods*; D014771:Virion / Q000302:isolation & purification*; D014776:Virus Cultivation
chemical_list: D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens