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Year : 2015  |  Volume : 33  |  Issue : 2  |  Page : 290-292

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


Kusuma School of Biological Sciences, Indian Institute of Technology Delhi, New Delhi, India

Correspondence Address:
P Vivekanandan
Kusuma School of Biological Sciences, Indian Institute of Technology Delhi, New Delhi
India
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Source of Support: DBT funded project (SAN.No.102/IFD/SAN/PR- 2454/2010-2013), Conflict of Interest: None


DOI: 10.4103/0255-0857.153568

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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.






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2004 - Indian Journal of Medical Microbiology
Published by Wolters Kluwer - Medknow

Online since April 2001, new site since 1st August '04