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目的构建基于纳米金探针和蛋白芯片的快速、多肿瘤标志物高灵敏检测体系。方法将待检测的肿瘤标志物捕获抗体(捕获待测抗原)结合在醛基化修饰的玻片上;用检测抗体制备纳米金探针;经过免疫反应形成三明治夹心结构(蛋白芯片-目标蛋白-纳米金探针)。利用纳米金沉积的方法进行染色,通过肉眼或显微镜观察显色结果。结果该蛋白检测体系在1 h内可检测两种肿瘤标志物,其中CEA可检测到最低浓度为45 pg/mL,CYFRA21-1可检测到90 pg/mL,与传统的酶联免疫吸附法(ELISA)相比,检测灵敏度大大提高。并利用此体系,检测肺癌患者及正常人血清,结果与临床所用电化学分析方法一致。结论该检测体系操作简单、方便、快速,同时具有高灵敏度、多指标联合检测等优点,在临床蛋白检测中具有重要的价值和应用前景。
OBJECTIVE To construct a rapid and multi-tumor marker highly sensitive detection system based on gold nanoprobe and protein chip. Methods Tumor markers to be detected capture antibody (capturing the antigen to be tested) binds to the aldehyde-modified glass slide; using the detection antibody to prepare gold nanoparticle probe; immunological reaction to form a sandwich sandwich structure (protein chip - target protein - nano Gold probe). Dyeing was carried out by means of deposition of gold nanoparticles, and the color development was observed by the naked eye or the microscope. Results The protein detection system could detect two kinds of tumor markers within 1 h, of which CEA could detect the lowest concentration of 45 pg / mL, CYFRA21-1 could detect 90 pg / mL, compared with the traditional enzyme-linked immunosorbent assay ELISA), the detection sensitivity is greatly improved. And the use of this system, detection of lung cancer patients and normal serum, the results with the clinical use of electrochemical analysis method. Conclusion The detection system is simple, convenient and rapid, and has the advantages of high sensitivity and multi-index joint detection. It has important value and application prospect in clinical protein detection.