甲胎蛋白双孔滴金免疫测定法试剂的研制及应用

应用自制的两种抗甲胎蛋白（ＡＦＰ）单克隆抗体Ａ_６Ｃ_６和Ｇ_４,分别包被硝酸纤维素膜及制备胶体金结合物,建立了检测ＡＦＰ的滴金免疫测定法。制备的试剂盒中应用双孔反应板,在相同条件下以ＡＦＰ参照标准（含ＡＦＰ５０μｇ／Ｌ）与血清标本作对比测定,可得到半定量结果,并实施试验的质量控制。试剂质量稳定,测定反应性良好。临床试用中肝细胞性肝癌的阳性检出率为９６．４％（２７／２８）。在与ＲＩＡ及ＥＩＡ的２０８例（包括其他疾病和健康人）对比测定中符合率分别为９９．１５％（１１６／１１７）和１００％（９１／９１）。此法快速简便,适合于各级医疗单位用作肝癌的实验室诊断方法。

Reagent Preparative by Two-well Dot Immunogold Filtration Assay for AFP Determination and Its Clinical Application

A dot immunogold filtration assay for the determination of Alpha-fetoprotein (AFP)in serum has been established by using two matched monoclonal antibodies (A_6C_6 and G_4) prepared in Our laboratory. A_6C_6 was inoculated on nitrocellulose menbrane, and G4 conjugated with colloidal gold. In the reagent kit prepared, a two-well device was specially designed to allow the assay performed with a reference standard as quality control, and a semi-quantitative result can be obtained. The reagent was stable, the assay was reproducible. In the clinical trail, a 96.4% positive rate was observed in 27/28 cases of hepatic carcinoma. In the comparative determination with RIA and EIA of 208 serum specimens including other disease and healthy person the correlation rate were 99.15% (116/117) and 100% (91/91) respectively. This assay is rapid and simple. It is suitable to be used in hospitals of different level as a laboratory diagnostic method for liver cancer