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本研究以时间分辨激发荧光光谱分析技术为基础 ,进行稀土离子标记的激光激发的时间分辨荧光免疫分析 (TRFIA)研究。实验以自行合成的二乙三胺五醋酸酐 (DTPAA)为双功能螯合剂。用 Eu3+标记兔抗人 (RAH) Ig G抗体 ,依据解离增强原理 (DEL FIA) ,研究了 Eu3+ -β萘甲酰三氟丙酮 (β- NTA)的荧光分辨体系 ,测定了荧光光谱和荧光寿命 ,建立了铕离子分析检出方法 ,其工作曲线范围为 1× 10 - 7～ 1× 10 - 1 1g· m L- 1 ,检测限为 1× 10 - 1 3g· m L- 1 ,相对标准偏差为 6 .4%。结合 TRFIA方法学研究 ,进行了人血清丙型肝炎病毒抗体 (Anti- HCV)检测。并同酶联免疫法 (EL ISA)对比。取得 TRFIA法阳性检测率明显高于EL ISA法的结果。
In this study, time-resolved fluorescence immunoassay (TRFIA) based on time-resolved excitation fluorescence spectroscopy was used to study the laser-induced fluorescence excitation of rare earth ions. Diethylenetriaminepentaacetic anhydride (DTPAA) was synthesized by ourselves as a bifunctional chelating agent. The Eu3 + -β-naphthoyltrifluoroacetone (β-NTA) fluorescence resolution system was studied by using Eu3 + labeled rabbit antihuman (RAH) Ig G antibody and the principle of dissociation enhancement (DEL FIA). The fluorescence spectra and fluorescence The working curve of Eu3 + ion was established. The working curve range was 1 × 10-7 ~ 1 × 10-1 1g · m L-1 with the detection limit of 1 × 10-1 3g · m L- 1, The standard deviation is 6.4%. Combined with TRFIA methodological studies, human serum anti-HCV antibody (Anti-HCV) was tested. And compared with enzyme-linked immunosorbent assay (ELISA). The positive rate of TRFIA was significantly higher than that of EL ISA.