Development of a time-resolved fluoroimmunoassay (TR-FIA) for testosterone: measurement of serum testosterone concentrations after testosterone treatment in the rainbow trout (Oncorhynchus mykiss).
Development of a time-resolved fluoroimmunoassay (TR-FIA) for testosterone: measurement of serum testosterone concentrations after testosterone treatment in the rainbow trout (Oncorhynchus mykiss).
复制标题
开发睾酮时间分辨荧光免疫测定法 (TR-FIA)：测量虹鳟 (Oncorhynchus mykiss) 睾酮治疗后的血清睾酮浓度。
DOI：
10.1006/gcen.1996.6861
复制
发表时间：
1997
影响因子：
2.7
通讯作者：
M. Iwata
中科院分区：
文献类型：
--
作者：
H. Yamada;R. Satoh;T. Yamashita;A. Kambegawa;M. Iwata
建立了一种高灵敏度的时间分辨荧光免疫分析法（TR-FIA），并将其应用于虹鳟鱼血清睾酮浓度的测定。用物理吸附法将四环素-3-（O-羧甲基）肟-牛血清白蛋白（T-3-CMO-BSA）固定在微孔板的威尔斯孔中。使用两种抗体在固相中的T-3-CMO-BSA、未知量的睾酮、睾酮抗体和铕标记的二抗之间进行竞争性测定，然后使用时间分辨荧光计（DELFIA系统）进行测量。TR-FIA的灵敏度为0.075 pg/50微升样本（1.5 pg/ml），测定系统的范围在1.5 pg/ml和25 ng/ml之间。睾酮TR-FIA的批内和批间变异系数均较低，分别为1.62和6.38%以及2.96和8.29%。该测定系统被应用于测量血清睾酮浓度后，注射睾酮溶解在盐水，丙二醇，或椰子油。在三种溶剂中，椰子油组显示持续高血清睾酮水平。相反，盐水和丙二醇组在注射后24小时具有最大浓度，但它们的水平显著低于椰子油组。睾酮TR-FIA方法是敏感的，可重复的，是准确的传统RIA。这是非常好的测量血清睾酮浓度。
A sensitive time-resolved fluoroimmunoassay (TR-FIA) for testosterone was developed, and the assay system was used for measuring serum testosterone concentrations in rainbow trout. Testosterone-3-(O-carboxymethyl)oxime-bovine serum albumin (T-3-CMO-BSA) was immobilized by physical adsorption to the wells of microtiter plates. A competitive assay using two antibodies was performed among T-3-CMO-BSA in the solid-phase, unknown amounts of testosterone, testosterone antibodies, and europium labeled secondary antibodies, followed by measurements using a time-resolved fluorometer (DELFIA system). The TR-FIA had a sensitivity of 0.075 pg/50 microliters sample (1.5 pg/ml), and the range of the assay system was between 1.5 pg/ml and 25 ng/ml. The intra- and interassay coefficients of variation for the testosterone TR-FIA were satisfactorily low, and were between 1.62 and 6.38% and 2.96 and 8.29%, respectively. The assay system was applied to measure the serum testosterone concentrations after an injection of testosterone dissolved in saline, propyleneglycol, or coconut oil. Among the three solvents, the coconut oil group showed continuously high serum testosterone level. In contrast, the saline and propyleneglycol groups had maximum concentrations 24 hr after the injection, but their levels were significantly lower than that of the coconut oil group. The testosterone TR-FIA method is sensitive, repeatable, and is as accurate as conventional RIAs. It is very good for measuring serum testosterone concentrations.