摘要
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。
This paper described a laser-excited time-re so lved fluoroimmunoassay set.It made lanthanide ion to couple the anhydrde of diet hylenetriaminepentaacetic acid (DTPAA) for labeling antibodies.The experiment us ed polystyrene tap coated with HCV antigen as the solid phase and a chelate of t he rare eart h metal europium as fluorescent label.A nitrogen laser beam was used to excite t he Eu- 3+ chelates and after 60 μs delay time,the emission fluorescence w as measured.Background fluorescence of short lifetimes caused by serum component s and Raman scattering can be eliminated by set the delay time.In the system con dition,fluorescent spectra and fluorescent lifetimes of Eu- 3+ β-nap hthoyltrifluroacetone (NTA) chelates were measured.The fluorescent lifetime valu e is 650 μs.The maximum emssion wavelength is 613 nm.The linear range of europi um ion concentration is 1×10- -7—1×10- -11 g·mL- -1 and the detection limit is 1×10- -13 g·mL- -1.The relative s tandard deviation of determination (n=12) for samples at 0.01 ng·mL- - 1 magnitude is 6.4%.Laser-TRFIA was also found to be suitable for diagnosis of HCV.The sensitvity and specificity were comparable to enzyme immunoassay.The result was obtained with laser-TRFIA for 29 human correlated well with enzyme immunoassay.
出处
《光谱学与光谱分析》
SCIE
EI
CAS
CSCD
北大核心
2000年第3期277-279,共3页
Spectroscopy and Spectral Analysis
基金
国家自然科学基金项目
