快速检测甲胎蛋白的免疫层析试条的研制

Study on the Preparation of Immunochromatographic Strip for Rapid Detection of α-Fetoprotein

研制了可简便、快速检出原发性肝癌标志物——人血清中甲胎蛋白 ( AFP)的免疫层析试条。为此 ,研究了用国产试剂和材料制备胶体金、用此胶体金标记抗 AFP单抗的影响因素以及抗 AFP抗体固化条件、抗 AFP抗体与其 AFP结合能力等。结果表明 ,自制国产胶体金质量不亚于进口商品 ;金标单抗和固化抗体活性稳定 ,能分别显示出结合 AFP分子不同表位的特异性。所得检测试条的检出速度在 5至 2 0 min内 ,检测阳性阈值定为 2 0 ng AFP/m L血清。

An immunochromatographic strip for the simple and rapid detection of α fetoprotein (AFP) in human serum, a marker for primary hepatic carcinoma is developed in this study. The preparation of colloidal gold was accomplished with the reagents and materials made in China. The factors influencing anti AFP monoclonal antibodies labeled with the colloidal gold were tested. The conditions for the immobilization of the anti AFP antibodies and the binding ability of the anti AFP antibodies with AFP were also investigated. The results revealed that the colloidal gold prepared by ourselves was as good as the commercial ones. The activities of the monoclonal antibodies labeled with the colloidal gold and the immobilized antibodies were stable. The monoclonal antibodies labeled with the colloidal gold and the immobilized antibodies showed specificities of binding different epitopes on AFP molecule. The time for the detection with the prepared strips was 5 to 20 min, and the cut off value for positive result was 20 ng AFP/mL of serum.