Lewis Mark G, Lewis John G, Elder Peter A, Moore Grant A
Canterbury Health Laboratories, Christchurch, New Zealand.
J Anal Toxicol. 2003 Sep;27(6):342-5. doi: 10.1093/jat/27.6.342.
A direct enyzme-linked immunosorbent assay (ELISA) for urinary immunoreactive methylphenidate (Ritalin), in which a standard 96-well microtiter plate is used, is described. For this ELISA, a methylphenidate-thyroglobulin conjugate is immobilized to the microtiter plate and competes with methylphenidate in the standard or urine sample for antibody-binding sites. After washing, the sheep methylphenidate antibody bound to immobilized methylphenidate is detected with peroxidase-labelled goat antisheep IgG. Following a further wash, tetramethylbenzidine is added, color is developed, and the plate is read at 450 nm on an ELISA plate reader. This method is unaffected by drugs of abuse and is suitable for routine use in the toxicology laboratory.
本文描述了一种用于检测尿液中免疫反应性哌醋甲酯(利他林)的直接酶联免疫吸附测定(ELISA)方法,该方法使用标准的96孔微量滴定板。在这种ELISA中,哌醋甲酯-甲状腺球蛋白偶联物固定在微量滴定板上,并与标准品或尿液样本中的哌醋甲酯竞争抗体结合位点。洗涤后,用辣根过氧化物酶标记的山羊抗绵羊IgG检测与固定化哌醋甲酯结合的绵羊哌醋甲酯抗体。进一步洗涤后,加入四甲基联苯胺,显色,然后在ELISA酶标仪上于450nm处读取吸光度。该方法不受滥用药物的影响,适用于毒理学实验室的常规检测。