Amplification of hepatitis B virus covalently closed circular DNA in patients with hepatocellular carcinoma using rolling circle amplification

Abstract:

Objective：To analyze the intrahepatic hepatitis B virus（HBV） covalently closed circular DNA（cccDNA） in hepatocellular carcinoma（HCC） patients. Methods：The specificity and sensitivity of rolling circle amplification（RCA） were investigated. The intra－hepatic HBV cccDNA in 13 pairs of tumor and adjacent non-tumor tissues were amplified by RCA and pre-S deletions were deter－mined by sequencing. Results：The HBV cccDNA（10 copy/μl） could be amplified specifically by RCA. Among 26 samples，the HBV cccDNA in 22 tissues were amplified successfully，and the pre-S deletions were identified in 8 samples（8/22，36.4%）. In addition，the rate of pre-S2 deletion was higher than that of pre-S1（8/8 vs. 2/8，P=0.007）. Conclusion：The pre-S2 deletion of HBV cccDNA is first identified in hepatocellular carcinoma patients，which would be benefit for understanding the relationship between cccDNA pre-S deletion and HCC.