流动注射化学发光同时检测甲胎蛋白和癌胚抗原

Chemiluminescence Immunoassay for Simultaneous Detection of α-Fetoprotein and Carcinoembryonic Antigen

将包被有单克隆抗体的两个透明微型检测池串联在流动注射分析装置上,利用移动式微型检测池建立了一种能同时检测甲胎蛋白和癌胚抗原的流动注射化学发光免疫分析方法,为提高分析效率提供了一种有效途径。样品抗原和辣根过氧化物酶标记的相应抗体分别导入微管进行孵育,形成三明治式免疫夹心结构。在注入鲁米诺和H2O2后,两微型检测池中分别形成较稳定的酶催化化学发光体系。通过切换检测池,使两个透明微型检测池中发生的酶促增强化学发光反应相继得到检测,从而实现两种待测物的同时检测。考察了一系列影响化学发光检测的参数,如免疫反应时间、鲁米诺和H2O2的浓度及反应介质的pH值。在最佳实验条件下,甲胎蛋白和癌胚抗原的检测线性范围分别为1.25～50.00μg/L和1.25～40.00μg/L,检出限分别为1.06和1.00μg/L。对人血清实际样品进行了检测,取得了满意的结果。

A novel flow-through chemiluminescent （CL） immunoassay was designed for the simultaneous detection of α-fetoprotein （AFP） and carcinoembryonic antigen （CEA）.Two transparent microtubes immobilized with monoclonal antibodies were set in a mobile flow cell.Based on a sandwich immunoassay format,two mixtures of the sample antigens and corresponding horseradish peroxidase （HRP） labeled antibodies were introduced into the channels for on-line incubation.After the injection of luminol and H2O2 solution,different CL signals from the two transparent microtubes were sequentially detected by moving the mobile flow cell.The effects on the chemiluminescence detection,such as the immunological reaction time,the concentrations of luminol and H2O2,and the pH value of the buffer solution,were carried out.Under the optimal experimental conditions,AFP and CEA could be detected in the linear ranges of 1.25-50 μg/L and 1.25-40.0 μg/L with detection limits of 1.06 and 1.0 μg/L,respectively.The actual samples of human serum was analyzed with satisfactory results.