A diagnostic method based on PCR for detection of hepatitis B virus X gene deletion mutant HBx-d382

Objective To develop PCR method for detection of HBV X gene deletion mutant HBx-d382 and study its clinical value in the diagnosis of HBV-related hepatocellular carcinoma. Methods Specific primers for HBx-d382 mutant were designed by bioinformatics, and PCR method was established to detect HBx gene and HBx-d382 mutant gene in 82 sera of patients with hepatocellular carcinoma, 92 sera of patients with chronic HBV infection and 20 sera from healthy controls. The HBx genes were cloned, sequenced and aligned with the published HBx gene sequence, and HBx gene PCR results were compared with HBx gene sequencing results. Results The detection rate of HBx-d382 was 2.1% in sera of patients with chronic HBV infection and 17.1% in sera of patients with hepatocellular carcinoma. Compared with chronic HBV infection,there was a higher HBx-d382 detection rate in sera of patients with hepatocellular carcinoma(χ2=4.21, P0.05), and HBx gene PCR results consistent with HBx gene sequencing results. Conclusion PCR method for detection of HBV X gene deletion mutants HBx-d382 cans be used for early diagnosis of HBV-related hepatocellular carcinoma.