Choi M J, Gorovits B M, Choi J, Song E Y, Nam K S, Park J
Doping Control Center, Korea Institute of Science and Technology, Seoul.
Biol Pharm Bull. 1994 Jul;17(7):875-80. doi: 10.1248/bpb.17.875.
A visual membrane enzyme immunoassay is described for the measurement of methamphetamine in urine. To increase assay sensitivity, tracers with chemically similar structures were cross-checked with the antibodies to determine their influence on the antibody binding. Tracers of horseradish peroxidase-labeled methamphetamine (MA-HRP) and amphetamine (A-HRP) derivatives were prepared for this purpose. Significant differences in antibody specificity were found between the two tracers. Based on the results of this study, a pair of an antibody and a tracer was selected and a membrane enzyme immunoassay (EIA) was developed utilizing the competitive binding between methamphetamine and the drug-HRP tracer. UltraBind membrane (0.45 micron) was used as the solid matrix to which the antibody was attached. Using diaminobenzidine substrate with Co2+ ion, a stable grey color appeared on the surface of membrane for MA-negative urine samples. No color appeared for MA-positive urine with a cut-off level of 0.8 ppm.
本文描述了一种用于检测尿液中甲基苯丙胺的可视化膜酶免疫测定法。为提高检测灵敏度，用化学结构相似的示踪剂与抗体进行交叉检测，以确定它们对抗体结合的影响。为此制备了辣根过氧化物酶标记的甲基苯丙胺（MA-HRP）和苯丙胺（A-HRP）衍生物示踪剂。发现两种示踪剂之间抗体特异性存在显著差异。基于本研究结果，选择了一对抗体和示踪剂，并利用甲基苯丙胺与药物-HRP示踪剂之间的竞争结合开发了一种膜酶免疫测定（EIA）。使用UltraBind膜（0.45微米）作为附着抗体的固体基质。对于甲基苯丙胺阴性尿液样本，使用含Co2+离子的二氨基联苯底物时，膜表面会出现稳定的灰色。对于甲基苯丙胺阳性尿液，当截断水平为0.8 ppm时无颜色出现。
