A visible assay for evaluating the inhibitory activity of drug and antibody against HBV infection

ABSTRACT Viral infectivity detection assays play an important role in the quantitative titers of virus and the effect of drugs and antibodies against virus infection. However, detecting the infectivity of hepatitis B virus (HBV) via conventional plaque assays still poses challenges because its infection does not result in cytopathic effects and progeny viruses struggle to infect adjacent cells in vitro . In this study, we established a sensitive and reliable immunospot assay for the visible quantitative detection of HBV infection based on an excellent detection antibody targeting the HBV core protein. Furthermore, this assay demonstrated robust performance in evaluating the inhibitory activity of antiviral drugs, as well as the neutralizing activity of monoclonal antibodies and plasma samples from vaccinated individuals against HBV infection. Collectively, our findings reported a sensitive and reliable method for the visible quantitative detection of HBV infectivity, offering significant potential for broad application in various HBV-related research fields. IMPORTANCE In this study, we report a sensitive and reliable immunospot assay for the visible quantitative detection of HBV infection. This assay demonstrates excellent performance in evaluating the inhibitory effect of antiviral drugs, neutralizing antibodies, and plasma samples from vaccinated individuals. Given its robustness and versatility, the proposed method is anticipated to be an excellent alternative assay for HBV-related research, including therapeutic development and vaccine evaluation.