甲胎蛋白和癌胚抗原的胶体金免疫层析同步检测

Study on Gold-immunochromatographic Assay for Simultaneous Detection of AFP and CEA

目的:探讨建立一种同时快速检测甲胎蛋白(AFP)和癌胚抗原(CEA)两种肿瘤标志物的方法。方法:用胶体金分别标记鼠抗AFP单克隆抗体和鼠抗CEA单克隆抗体,喷于胶金垫上,与之配对的抗体喷在硝酸纤维素膜(NC膜)上分别做检测线T2和T1,兔抗鼠二抗喷在NC膜上做质控线,利用免疫层析技术进行检测。结果:采用Frens方法制备的胶体金粒径在20 nm,紫外可见吸收峰在521 nm处,金标抗体的最适pH在9.0,最适抗体量为200μl,胶体金标记2.4μg抗体。测试结果表明同时检测AFP和CEA的试纸条灵敏度分别为20 ng.mL-1和5 ng.mL-1,检测时间仅需10 min,与PSA、CA125、CA15-3、Ferritin、Human Albumin等没有交叉反应,稳定性好。结论:应用胶体金免疫层析技术,实现了AFP和CEA的同步检测。

Objective： To establish a rapid assay for simultaneous detection of AFP and CEA with colloidal gold immunochro-matographic （ICG） technology. Methods： Colloidal gold was used to mark the anti-mouse monoclonal antibody against AFP and CEA, and then sprayed on conjugate pad of the test strip. The other two antibodies matched with AFP and CEA were sprayed on Nitrocellulose Membrane （NC membrane） as test line T2 and T1, and rabbit anti-mouse antibody was sprayed on NC membrane as control. Immunoehro-matographic technology has been used in the detection. Results： Colliodal gold prepared by Frens method was 20 nm, UV-visible absorption peak at 521 rim, optimal marked conditions of pH was 9.0, the optimal amount of antibody was 2.4μg antibody each 200μl colloidal gold. Results showed that the sensitive standard of rapid assay for AFP and CEA visual detection limit were 20 ng.mL^-1 and 5 ng. mL^-1, which only required 10 min. There was no cross-reaction with PSA, CA125, CA15-3, Ferritin, Human Albumin, and the rest strip was stable. Conclusions： AFP and CEA can be detected simultaneously with ICG technology.