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目的:建立双抗体间接夹心酶联免疫吸附法(ELISA)试剂盒,检测血清黏蛋白1(MUC1)水平,探讨其在肺癌诊断中的应用,阐明血清MUC1蛋白检测的临床应用价值。方法:在前期制备的MUC1蛋白和兔抗人MUC1多克隆抗体的基础上,建立以鼠抗人MUC1单克隆抗体为包被抗体,兔抗人MUC1多克隆抗体为检测抗体的双抗体间接夹心ELISA方法,并通过对MUC1蛋白标准品的检测,绘制出标准曲线。临床上收集48例肺癌患者、7例肺良性疾病患者和20例健康人的血清,应用本研究建立的ELISA方法检测各组标本血清MUC1蛋白表达水平,绘制ROC曲线,确定最佳cut-off值,得出双抗体间接夹心ELISA方法检测肺癌的敏感度、特异度及约登指数。同时与临床应用的CA15-3试剂盒检测结果进行比较。结果:应用双抗体间接夹心ELISA法确定血清MUC1的临界值为1.98μg.L-1,检测肺癌的敏感度为62.5%,特异度为100%,约登指数为0.625 0。CA15-3试剂盒检测肺癌的敏感度为18.75%,特异度为100%,约登指数为0.187 5。结论:建立的双抗体间接夹心ELISA试剂盒检测肺癌有更高的敏感度和特异度,优于临床常用的CA15-3试剂盒。
OBJECTIVE: To establish a sandwich ELISA kit for the detection of serum mucin 1 (MUC1) in lung cancer and to explore its clinical value in the diagnosis of lung cancer and elucidate the clinical value of detecting serum MUC1 protein. Methods: Based on the pre-prepared MUC1 protein and rabbit anti-human MUC1 polyclonal antibody, an indirect sandwich ELISA with the anti-human MUC1 monoclonal antibody as the coating antibody and the rabbit anti-human MUC1 polyclonal antibody as the detection antibody Method, and through the detection of MUC1 protein standard, draw a standard curve. Serum of 48 patients with lung cancer, 7 patients with benign lung disease and 20 healthy people were collected clinically. The serum MUC1 protein level in each group was detected by ELISA. The best cut-off value , The indirect antibody sandwich ELISA was used to detect the sensitivity, specificity and Youden index of lung cancer. At the same time with the clinical application of CA15-3 kit test results were compared. Results: The critical value of serum MUC1 was 1.98μg.L-1 by indirect antibody sandwich ELISA. The sensitivity and specificity of detection of MUC1 in serum were 62.5%, 100% and 0.6250, respectively. The sensitivity of CA15-3 kit for detecting lung cancer was 18.75%, the specificity was 100%, and the Youden index was 0.187 5. Conclusion: The established indirect sandwich ELISA kit for detecting lung cancer has higher sensitivity and specificity than the commonly used clinical CA15-3 kit.