Morra di Cella Stefania, Veglio Franco, Mulatero Paolo, Christensen Valerie, Aycock Keli, Zhu Zheng, Gomez-Sanchez Elise P, Gomez-Sanchez Celso E
Department of Internal Medicine, University of Torino, Italy.
J Steroid Biochem Mol Biol. 2002 Sep;82(1):83-8. doi: 10.1016/s0960-0760(02)00142-5.
Patients with primary aldosteronism and with glucocorticoid-suppressible aldosteronism excrete in the urine excessive amounts of the hybrid steroids 18-hydroxycortisol and 18-oxocortisol. The measurement of these steroids aids in the differential diagnosis of various adrenal disorders. We have produced mouse monoclonal antibodies against 18-oxocortisol and polyclonal antibodies against 18-hydroxycortisol and describe a time-resolved fluoroimmunoassay (TR-FIA) technique for the measurement of these steroids in the urine. We have also compared this assay with an ELISA technique for these compounds. We also describe the preparation of in-house Eu(III)-labeled avidin and an enhancement solution and compared to a commercially available Eu(III)-labeled streptavidin and enhancement solutions. The monoclonal antibodies against 18-oxocortisol are sensitive and have a high level of specificity. The TR-FIA technique using in-house prepared reagents or commercial ones were indistinguishable from each other, but at a significant saving. The TR-FIA technique was more sensitive and had a greater precision than the ELISA technique for both steroids.
原发性醛固酮增多症和糖皮质激素可抑制性醛固酮增多症患者尿中排泄过量的混合类固醇18-羟皮质醇和18-氧代皮质醇。这些类固醇的测定有助于各种肾上腺疾病的鉴别诊断。我们制备了抗18-氧代皮质醇的小鼠单克隆抗体和抗18-羟皮质醇的多克隆抗体，并描述了一种用于测定尿中这些类固醇的时间分辨荧光免疫分析（TR-FIA）技术。我们还将该分析方法与针对这些化合物的酶联免疫吸附测定（ELISA）技术进行了比较。我们还描述了自制铕（III）标记抗生物素蛋白和增强溶液的制备方法，并与市售铕（III）标记链霉抗生物素蛋白和增强溶液进行了比较。抗18-氧代皮质醇的单克隆抗体灵敏且具有高度特异性。使用自制试剂或市售试剂的TR-FIA技术彼此无明显差异，但成本大幅降低。对于这两种类固醇，TR-FIA技术比ELISA技术更灵敏且精密度更高。
