Hepatitis B Virus Serological and Viral Load Analysis

The serum of mice was collected and diluted 1:10 with PBS. HBsAg, HBsAb, HBeAg, HBeAb, and HBcAb were detected using an ELISA kit (Kehua Bio-engineering Co. Ltd., Shanghai, China), per the manufacturer's instructions. The viral load was quantified by real-time polymerase chain reaction (PCR) using SYBR Green dye (Sigma-Aldrich, St. Louis, MO, USA) as described previously (Wang et al., 2014b (link)). HBcAg in the liver tissue was detected by immunohistochemistry. The liver tissue was collected, embedded in paraffin, and sectioned. The sections were stained with rabbit anti- HBcAg polyclonal antibody (Dako, Japan) and visualized using the DAKO EnVision™ Detection Systems (Dako, Japan), according to the manufacturer's instructions.