Simultaneous detection of tumor markers in human serum using time-resolved fluorescence microspheres immunochromatographic assay / 中华核医学与分子影像杂志

ABSTRACT

Objective To develop a time-resolved fluorescent microspheres immunochromatographic assay (TRFMIA) for detection of alphafetoprotein (AFP) and carcinoembryonic antigen (CEA) in human serum and to evaluate its performance.Methods The Eu-time-resolved fluorescent polystyrene particles conjugated with monoclonal antibody AC18# for AFP and AE03# for CEA were used as fluorescent labels.The monoclonal antibody AC17# for AFP,AE05# for CEA and goat anti-rabbit antibody were immobilized on the nitrocellulose membrane as the test lines and control line.Several performances indicators were measured,including linear range,detection limit,and specificity.AFP and CEA were measured by the new method and the results were compared with those obtained by time-resolved fluoroimmunoassay (TRFIA) and electro-chemiluminescence immunoassay (ECLIA) using linear correlation analysis.Results The measurement ranges of AFP were 0.07-1 000.00 kU/L with the intra-and inter-assay CV of 5.93％ and 11.07％,and those of CEA were 0.12-500.O0 μg/L with the intra-and inter-assay CV of 7.53％ and 12.13％ respectively.The average recovery rate of AFP and CEA was 92.77％ and 94.73％,respectively.Measurements obtained by TRFMIA had strong correlation coefficients ranging from 0.93 to 0.97 when compared with those obtained by TRFIA and ECLIA.Conclusion TRFMIA,which can simultaneously detect AFP and CEA,has been successfully established.