Tresguerres J A, Lisboa B P, Tamm J
Steroids. 1976 Jul;28(1):13-23. doi: 10.1016/0039-128x(76)90121-5.
A simple, reliable and rapid radioimmunoassay (RIA) for the determination of testosterone glucosiduronate (TG) in crude urine is described. Two protein-TG complexes were investigated in raising antibodies: a) Bovine serum albumin (BSA)-TG and b) human plasma Cohn's fraction IV-4 (CF)-TG. In rabbits, high titers of antibodies were obtained after the injection of CF-TG. The specificity of the antiserum was sufficiently high (cross reaction with free testosterone 27%, with 5alpha-dihydrotestosterone-glucosiduronate 20%). TG was estimated in small aliquots of male and female urine after evaporation overnight at 50 degrees C in order to eliminate interfering material. The intraassay coefficient of variation (CV) was found to be 6% and the interassay CV 11%. TB has been determined in 40 samples of urine simultaneously by "direct" RIA and by a "classical" RIA following hydrolysis with beta-glucuronidase. The coefficient of correlation was found to be 0.89. The mean excretion of TG in the urine of 26 healthy men amounted to 164+/-51 mug/24 hours with a range from 97 to 346 mug/24 hours. In a group of 16 women a mean urinary excretion of TG of 24+/-10 mug/24 hours was determined. The method allows a technician to assay 40 samples per day.
本文描述了一种用于测定尿样中睾酮葡萄糖醛酸苷（TG）的简单、可靠且快速的放射免疫分析方法（RIA）。在制备抗体时研究了两种蛋白质-TG复合物：a）牛血清白蛋白（BSA）-TG和b）人血浆Cohn's 组分IV-4（CF）-TG。在兔子中，注射CF-TG后获得了高滴度的抗体。抗血清的特异性足够高（与游离睾酮的交叉反应为27%，与5α-二氢睾酮葡萄糖醛酸苷的交叉反应为20%）。为消除干扰物质，将男性和女性尿样的小份试样在50℃下过夜蒸发后，对TG进行测定。批内变异系数（CV）为6%，批间CV为11%。通过“直接”RIA和用β-葡萄糖醛酸酶水解后的“经典”RIA同时测定了40份尿样中的TG。发现相关系数为0.89。26名健康男性尿中TG的平均排泄量为164±51μg/24小时，范围为97至346μg/24小时。在一组16名女性中，测定的TG平均尿排泄量为24±10μg/24小时。该方法允许技术人员每天检测40份样品。
