HBeAg, HBsAg and anti-HBs were quantified using the Architect™ platform (Abbott Laboratories, Wiesbaden, Germany), anti-HBc using Enzygnost® Anti-HBc monoclonal test (Siemens Healthcare Diagnostics, Erlangen, Germany). Alanine aminotransferase (ALT) activity was determined after 1:4 dilution in PBS by Reflotron® GPT/ALT tests (Roche Diagnostics, Mannheim, Germany).
DNA was extracted from 50 μl mouse serum using the QIAamp MinElute Virus Spin Kit (Qiagen, Hilden, Germany) or 20 mg of liver tissue using a NucleoSpin Tissue DNA Kit (Macherey-Nagel, Dueren, Germany) according to the manufacturer’s instructions. The quantification of HBV-DNA was performed through real-time PCR with SYBR green as previously described.22
DNA was extracted from 50 μl mouse serum using the QIAamp MinElute Virus Spin Kit (Qiagen, Hilden, Germany) or 20 mg of liver tissue using a NucleoSpin Tissue DNA Kit (Macherey-Nagel, Dueren, Germany) according to the manufacturer’s instructions. The quantification of HBV-DNA was performed through real-time PCR with SYBR green as previously described.22