乙肝病毒突变体HBx-d382 PCR检测方法的建立及临床应用

目的建立检测乙肝病毒突变体HBx-d382的PCR方法,探讨HBx-d382突变体在诊断HBV相关肝细胞癌中的应用价值。方法运用生物信息学和核苷酸合成的方法,设计和合成检测乙肝病毒突变体HBx-d382的特异性引物。采用PCR方法对82份HBV相关肝细胞癌病人血清、96份慢性HBV感染者血清和20份正常对照者血清中HBx基因进行检测。采用基因测序法对HBx基因进行分析,并与PCR结果进行比较。结果在慢性HBV感染者和肝细胞癌病人血清中,HBx-d382的检出率分别为2.1%与17.1%。与慢性HBV感染者相比,肝细胞癌病人血清中HBx-d382检出率较高(χ2=4.21,P0.05),且PCR检测结果与基因测序结果相符。结论肝细胞癌病人血清中存在乙肝病毒突变体HBx-d382,采用PCR方法检测HBx-d382突变体对早期诊断HBV相关肝细胞癌有一定的价值。

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 bioirfformatics, 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 （X2=4.21, P〈0.05）, 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 hepatocetlular carcinoma.