磁免疫电化学发光检测肺癌血清p53抗体

Immunomagnetic-electrochemiluminescent Detection of p53 Antibodies in Human Lung Cancer Serum

肿瘤抑制基因———p53基因的突变可能产生p53抗体 .p53抗体在肿瘤的诊断、预后及监测等方面具有重要的临床价值 .目前检测 p53抗体的方法 ,如酶联免疫分析方法 ,需要多个步骤 ,比较费时 ,且大部分检测指标只能是半定量 ,具有一定的局限性 .提出了一种磁免疫电化学发光 (IM ECL)分析方法定量检测人血清p53抗体 .这种新的分析方法检测人血清 p53抗体的最低检测极限可达到 10ng/L ,标准曲线的动力学范围和线性范围达到 5个数量级 (0 0 1～ 10 0 0 μg/L) .我们应用IM ECL分析法检测肺癌病人血清 ,只需要 50 μl的样品量 ,3 0min的孵育时间和少于 50s的采集时间 ,得出肺癌血清中p53抗体的阳性率为 2 8 6% ,然后通过标准曲线定量阳性血清中 p53抗体的浓度 .从肺癌血清的结果中发现 ,随着临床分期的升高 ,p53抗体浓度增加 .IM ECL分析方法在检测限、线性范围、分析时间等方面都优于酶联免疫分析 ,是一种可行的快速、灵敏、定量检测人血清

The mutation of tumor suppressor p53 gene probably produce p53 antibodies. p53 antibodies are of importance to tumor's diagnosis, prognosis and relapse monitor. Current method to detect p53 antibodies, i.e. enzyme-linked immunosorbent assay ( ELISA), requires long time with multi-step, and the assay is only semi-quantitative. Immunomagnetic electrochemiluminescence (IM-ECL) assay was devoloped for quantitative detection of p53 antibodies in human serum. Results indicate that the detection limit of this assay is 10 ng/L. A stable calibration curve with a wide linear range from 0.01 to 1 000 mug/L was established, thus, made quantitation possible. Serum samples from lung cancer patients were tested using the IM-ECL assay. The assay uses only 50 mul of sample per test and requires a 30 min incubation period in addition to a 50 s acquisition time. The positive rate of p53 antibodies was 28.6% in lung cancer sera. Concentrations of p53 antibody in the positive carcinoma sera were quantified from the calibration curve. A trend was found that higher the p53 antibody concentration in the lung cancer serum likely linked to a higher diagonostic stage of the cancer. IM-ECL assay is significantly better in terms of detection limit, linear range, and assay time, compared to the similar paramenters of common ELISA. The results suggests that IM-ECL is a feasible method for rapid, sensitive and quantitative detection of p53 antibodies in human serum.