Quantitative measurement of hepatitis B virus cccDNA

摘要

Hepatitis B virus (HBV) infection results in the entry of viral genomic DNA into host hepatocytes. This viral relaxed circular DNA (rcDNA) is transported to the nucleus, converted to covalently closed circular DNA (cccDNA), and serves as a template for viral transcription. Elimination of cccDNA is required to treat HBV infection, which remains a major therapeutic challenge. The robust and sensitive method for measuring cccDNA described herein is useful for promoting drug development and monitoring the effectiveness of the treatment. A set of primers was designed in combination with sodium bisulfite treatment of viral DNA to allow specific amplification of cccDNA without interference with the amplification of rcDNA. This method can be used to further guide therapeutic development and provide a non-invasive alternative for monitoring HBV-infected patients undergoing antiviral therapy.