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目的自制高尔基蛋白73(GP73)单克隆抗体并检测其诊断肝细胞肝癌(HCC)的敏感性和特异性。方法自行制备GP73单克隆抗体,作为一抗用Western blot方法检测59例HCC患者和31名健康对照者血清中GP73蛋白的表达情况,并对结果进行半定量分析,确定对照组人群的正常基线水平。将此结果同多克隆抗体的检测结果及甲胎蛋白(AFP)的诊断结果进行比较。结果使用单克隆抗体检测健康人群的血清GP73水平为1.2(0.9-1.7)相对单位(RU),HCC患者血清GP73水平为5.7(2.5-7.8)RU,显著高于对照组水平(P<0.001);多克隆抗体检测结果显示,HCC患者血清GP73水平与对照组相比[7.8(3.0-12.4)RU比1.1(1.0-2.0)RU]亦显著增高(P<0.001)。GP73单抗诊断HCC的敏感性和特异性分别为84.7%和93.5%,对比多抗的敏感性(78.0%)和特异性(93.5%),前者敏感性有所增高;同组患者采用AFP诊断HCC的敏感性和特异性分别为67.8%和74.2%,均低于GP73单抗。Logistic回归分析显示,GP73单抗的优势比(OR)为7.18,而GP73多抗的OR为1.51。结论单克隆抗体可以有效地检测肝癌患者血清GP73水平,比AFP具有更高的敏感性和特异性;可能优于目前使用的GP73多克隆抗体的检测。这一结果为用自制单抗进一步开发酶联免疫方法检测奠定了基础。
Objective To make the monoclonal antibody against GP73 and to test its sensitivity and specificity in diagnosing hepatocellular carcinoma (HCC). Methods GP73 monoclonal antibody was prepared by ourselves and used as a primary antibody to detect the expression of GP73 protein in 59 HCC patients and 31 healthy controls by Western blot. The results were semi-quantitatively analyzed to determine the normal baseline level of the control group . The results were compared with the results of the polyclonal antibody test and the diagnosis of AFP (AFP). Results Serum levels of GP73 in healthy individuals were 1.2 (0.9-1.7) relative units (RU) using monoclonal antibodies and 5.7 (2.5-7.8) RU in HCC patients, which was significantly higher than that of the control group (P <0.001) . The results of polyclonal antibody test showed that the serum GP73 level of HCC patients was significantly higher than that of the control group [7.8 (3.0-12.4) RU vs 1.1 (1.0-2.0) RU] (P <0.001). The sensitivities and specificities of GP73 monoclonal antibody in the diagnosis of HCC were 84.7% and 93.5%, respectively. The sensitivity and specificity of polyclonal antibody against GP73 were higher (78.0% and 93.5% The sensitivity and specificity of HCC were 67.8% and 74.2%, respectively, which were lower than that of GP73 monoclonal antibody. Logistic regression analysis showed that the odds ratio (OR) of GP73 McAb was 7.18, while the OR of GP73 polyclonal antibody was 1.51. Conclusion Monoclonal antibody can detect GP73 in patients with hepatocellular carcinoma effectively, which is more sensitive and specific than that of AFP. It may be superior to the currently used GP73 polyclonal antibody. This result laid the foundation for the further development of enzyme-linked immunosorbent assay using homemade monoclonal antibodies.