神经元特异性烯醇酶和癌胚抗原胶体金免疫层析联合检测

目的:应用双抗夹心胶体金免疫层析方法,实现对神经元特异性烯醇化酶(NSE)和癌胚抗原(CEA)两种肺癌肿瘤标志物的快速联合检测。方法:采用柠檬酸三钠还原法制备20nm胶体金颗粒,并分别对鼠抗NSE、CEA单克隆抗体进行标记,分别与之相配对的另一种单克隆抗体被喷在硝酸纤维素膜(NC膜)上,制成免疫层析检测试条。溶液中的抗原NSE、CEA与金标记抗体结合后沿着硝酸纤维素膜移动,与膜上固定的抗体结合形成肉眼可见的红色线条。结果:该试纸条只与NSE、CEA有特异性反应,与CA125、CYFRA21-1、TPA等肺癌标志物无交叉反应。标准样品中两种抗原的检测灵敏度分别可达到5ng/mL和3ng/mL。结论:胶体金免疫层析技术检测NSE、CEA特异性强、灵敏度高、简便快速,不需特殊仪器设备,有广泛应用价值。

Joint Detection of Neuron-specific Enolasc and Carcinoembryonic Antigen with Gold Immunochromatography Assay(GICA)

Objective: To realize the rapid joint detection for Neuron-specific enolase and carcinoembryonic antigen by double antibodies sandwich gold immunochromatography assay. Methods: Colloidal gold with diameter 20nm was prepared through reducing chloroauric acid with trisodium citrate. Gold nanoparticles were coupled with monoclonal anti-NSE/CEA. Another corresponding monoclonal antibody to the two antigens was fixed on the NC membrane to prepare GICA test strips. The gold-anti-NSE/CEA conjugate reacts with NSE/CEA in solution and migrates on the nitrocellulose membrane on which anti-NSE/CEA was immobilized, and producd the visual red lines. Results: This kind of test strips could react with NSE and CEA specificly, and there were no cross-reactions with other lung cancer markers, such as CA125, TPA, CYFRA21-1. The sensitivity of NSE was 5ng/mL and CEA was 3ng/mL by detecting the standard samples. Conclusion: GICA is a sensitive, specific,simple and rapid assay for detecting NSE and CEA. It need not any special devices and has significant practical value.