Elder P A, Lewis J G
J Steroid Biochem. 1985 May;22(5):635-8. doi: 10.1016/0022-4731(85)90217-1.
A rapid, single extraction ELISA for testosterone in plasma is described, using a standard 96 well microtitre plate. Testosterone is covalently bonded to bovine thyroglobulin and passively adsorbed in guanidine hydrochloride to the ELISA plate, giving an immobilised antigen approach which simplifies subsequent assay standardisation for steroid hormone assays. The addition of standard, sample and first antibody (rabbit anti-testosterone), which is unique for each different assay, is followed by a general procedure which includes washing, addition of peroxidase labelled goat antirabbit IgG, further washing and finally, addition of o-phenylenediamine substrate with colour development and reading of the plate at 492 nm on an automatic ELISA processor. The ELISA assay is compared to a testosterone RIA with 125I-label and has similar specificity and precision to the latter with a quicker processing time, and is more cost effective. The added advantages that ELISA assays confer over RIAs in terms of isotope purchase and disposal make this an ideal procedure for use in a routine steroid laboratory.
本文描述了一种用于血浆中睾酮的快速单提取酶联免疫吸附测定（ELISA）方法，该方法使用标准的96孔微量滴定板。睾酮与牛甲状腺球蛋白共价结合，并通过盐酸胍被动吸附到ELISA板上，形成一种固定化抗原方法，简化了随后甾体激素测定的标准化流程。加入标准品、样品和每种不同测定特有的第一抗体（兔抗睾酮抗体）后，按照通用程序进行操作，包括洗涤、加入过氧化物酶标记的山羊抗兔IgG、进一步洗涤，最后加入邻苯二胺底物显色，并在自动ELISA分析仪上于492nm处读取板值。将该ELISA测定法与125I标记的睾酮放射免疫测定（RIA）进行比较，结果显示该方法与后者具有相似的特异性和精密度，且处理时间更短，成本效益更高。ELISA测定法在同位素购买和处置方面优于RIA的额外优势，使其成为常规甾体实验室的理想检测方法。
