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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,P<0.05),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。
Objective To establish a PCR method for detecting hepatitis B virus (HBV) mutant HBx-d382 and to explore the value of HBx-d382 mutant in the diagnosis of HBV-related hepatocellular carcinoma. Methods The bioinformatics and nucleotide synthesis methods were used to design and synthesize specific primers for HBx-d382 detection. HBx gene was detected by PCR in sera from 82 HBV-related hepatocellular carcinoma patients, 96 serum from chronic HBV infected patients and 20 normal controls. HBx gene was analyzed by gene sequencing and compared with PCR results. Results The detection rates of HBx-d382 in patients with chronic HBV infection and those with hepatocellular carcinoma were 2.1% and 17.1%, respectively. HBx-d382 was higher in patients with hepatocellular carcinoma than in patients with chronic HBV infection (χ2 = 4.21, P <0.05), and the PCR results were in good agreement with those of the gene sequencing. Conclusion HBx-d382 is present in the serum of patients with hepatocellular carcinoma. The detection of HBx-d382 by PCR has some value in the early diagnosis of HBV-related hepatocellular carcinoma.