乙型肝炎病毒X基因的原核表达及血清抗HBx抗体的检测

目的 应用表达蛋白检测乙型肝炎患者血清抗-HBx抗体水平并探讨其临床意义。方法 通过PCR扩增获得HBVX基因，与原核表达载体Pet32a＋连接构建PET32a-HBX原核表达载体，转化E．coli BL21表达获得重组融合蛋白。经切胶透析纯化后，应用重组蛋白HBx建立检测血清中抗-HBx抗体的间接ELISA方法，分别检测正常人组、急性肝炎组、慢性肝炎组、肝硬化组和肝细胞癌组患者血清中的抗-HBx抗体。结果 获得具有免疫原性的HBx融合蛋白；ELISA检测表明，慢性肝炎组、肝硬化组和肝细胞癌组的抗-HBx抗体的水平均高于急性肝炎组，差异具有显著性；在三组之问，慢性肝炎组高于肝硬化组和肝细胞癌组，差异具有显著性，肝硬化组和肝细胞癌组的抗-HBx抗体水平无显著性差异。结论 HBV患者血清中抗-HBX抗体是乙型肝炎病毒感染的一种特异性抗体，是HBV感染的血清学指标之一，可以反映乙型肝炎肝炎患者病情的变化。

Erokaryotic expression and clinical application of X gene of hepatitis B virus

Objective To detect anti-HBx levels in serum of hepatitis B patients and study its clinical value. Methods The HBV X gene was amplified by PCR, ligated with the prokaryotic expression vector Pet32a , and transformed into E.coli BL21. The expressed fusion protein was identified by Western bloting. The anti-HBx of hepatitis B patients including NH (normal hedth) group, AH (acnte hepatitis) group, CH (chronic hepatitis) group, LC (liver cirrchosis) group and HCC group were detected by ELISA.Results Western blot analysis showed that the fusion HBx protein had good reactivity to the HBV positive serum. The level of anti-HBx of CH, LC and HCC groups were higher than those of AH group, the level of anti-HBx of CH group was higher than those of LC and HCC groups, there were no statistical difference between LC and HCC groups.Conclusion Anti-HBx antibody is one of the specific antibody and serological markers in hepatitis B patients, and the level of Anti-HBx antibody might reflect the state of illness of patients.