Barnard G, Kohen F, Mikola H, Lovgren T
Department of Hormone Research, Weizmann Institute of Science, Rehovot, Israel.
J Biolumin Chemilumin. 1989 Jul;4(1):177-84. doi: 10.1002/bio.1170040126.
We describe the principles of a new generation of sequential or simultaneous time-resolved fluoroimmunoassays, namely, simple, rapid, liquid-phase non-separation procedures which may be applied to the measurement of urinary steroid and drug metabolites. As an example, a method for the measurement of estrone-3-glucuronide in undiluted urine is reported. This method has a similar sensitivity, specificity and accuracy to a conventional separation fluoroimmunoassay or radioimmunoassay but in terms of speed, convenience, precision, reliability and clinical utility the new method has many advantages. The labelled antigen is a novel fluorescent europium chelate covalently linked to estrone-3-glucuronide. The antibody-binding reaction involves the incubation of the labelled antigen (2ng) with a limited concentration of polyclonal or monoclonal antibodies to estrone-3-glucuronyl-6-BSA and an aliquot of standard or sample (undiluted urine; 10 microliters) in microtitre wells. After a 10 min incubation, the fluorescence which emanates from the antibody-free label is measured in a time-resolved fluorometer and is proportional to the concentration of estrone-3-glucuronide in the standard or sample. The method may be applied for the monitoring of ovarian function in women.
我们描述了新一代顺序或同时时间分辨荧光免疫分析的原理，即简单、快速的液相非分离程序，可用于测量尿甾体和药物代谢物。作为一个例子，报道了一种测量未稀释尿液中雌酮 - 3 - 葡萄糖醛酸苷的方法。该方法与传统的分离荧光免疫分析或放射免疫分析具有相似的灵敏度、特异性和准确性，但在速度、便利性、精密度、可靠性和临床实用性方面，新方法具有许多优点。标记抗原是一种与雌酮 - 3 - 葡萄糖醛酸苷共价连接的新型荧光铕螯合物。抗体结合反应包括在微量滴定孔中，将标记抗原（2ng）与有限浓度的抗雌酮 - 3 - 葡萄糖醛酸 - 6 - BSA多克隆或单克隆抗体以及一份标准品或样品（未稀释尿液；10微升）一起孵育。孵育10分钟后，在时间分辨荧光计中测量来自无抗体标记物发出的荧光，其与标准品或样品中雌酮 - 3 - 葡萄糖醛酸苷的浓度成正比。该方法可用于监测女性的卵巢功能。
