联合检测甲胎蛋白和癌胚抗原的胶体金免疫层析试剂的研制及性能评价

Development of a gold immunochromatographic assay reagent for rapid detection of alpha-fetoprotein and carcinoembryonic antigen and evaluation of its performance

目的建立一种新型的联合检测甲胎蛋白(AFP)和癌胚抗原(CEA)的胶体金免疫层析法(G ICA)。方法G4和C2为一对抗AFP单克隆抗体(M cAb),A1和C9为另一对抗CEA M cAb,G4和A1分别以直线形式平行包被于固相硝酸纤维素(NC)膜上,C2和C9分别与胶体金结合后固定于结合物垫,制备成G ICA试剂条。膜上另包被2 mg/mL兔抗鼠IgG抗体,用于检测有效性。结果利用双抗体夹心原理,建立了可同时检测血清中AFP和CEA的G ICA。本法经AFP和CEA标准液测定,试剂的敏感性为AFP 30μg/L、CEA 10μg/L,与血红蛋白、人白蛋白、人球蛋白等无交叉反应,稳定性试验表明试剂可以在室温保存6个月以上。86份经化学发光免疫分析仪定量测定AFP、CEA浓度的临床血清标本用于本法的对比测定,符合性良好。结论联合检测AFP和CEA的G ICA试剂具有结果可靠、方法简便、快速等优点,与化学发光法测定结果符合性良好。

Objective To develop a novel gold immunochromatographic assay（GICA） for the detection of alpha-fetoprotein（AFP） and carcinoembryonic antigen（CEA） in serum.Methods The GICA strip was developed with one pair of anti-AFP monoclonal antibodies（McAb） G4/C2 and another pair of anti-CEA McAb A1/C9.G4 and A1 were coated on the nitrocellulose（NC） membrane as parallel lines,and C2 and C9 were labeled with colloidal gold respectively.2 mg/mL of rabbit anti-mouse IgG were also immobilized on the NC membrane for the controls of the assay.Results According to double antibody sandwich technique,the determination of AFP and CEA standard solution showed that the result of GICA could determine AFP and CEA levels simultaneously.The sensitivity of the kit was 30 and 10 μg/L in the determination of AFP and CEA respectively.The reagent was not interfered by hemoglobin,human albumin and human immunoglobulin and could be preserved at ambient temperature for 6 months.Simultaneous determination of 86 clinical serum samples by quantitative chemiluminescence immunoassay system and GICA gave coincident results.Conclusions The developed GICA for AFP and CEA is simple,rapid and reliable for clinical use.The strip determination result corresponds to those of chemiluminescence immunoassay.