24h尿液18-羟皮质醇ABC-ELISA的建立  

Development of A Competitive Avidin-Biotin Complex Enzyme Linked-Immunoassay for 18-Hydroxycortisol in Urine

作　　者：刘礼斌[1] 丁伟[1] 罗伟[2] 宓志均[1] 许曼音[1] 陈家伦[1] 

机构地区：[1]上海第二医科大学瑞金医院上海市内分泌研究所,上海200025 [2]上海第二医科大学瑞金医院检验重点实验室

出　　处：《上海第二医科大学学报》2002年第2期101-103,114,共4页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：国家自然科学基金资助课题 (3 95 70 3 45 )

摘　　要：目的建立 2 4h尿液 18-羟皮质醇亲合素 -生物素酶联免疫测定法 (ABC -ELISA)。　方法制备18-羟皮质醇抗血清 ,以此建立 18-羟皮质醇竞争性ABC -ELISA ;同时测定部分正常人、原醛腺瘤、原醛特发性增生病人 2 4h尿液 18-羟皮质醇水平。　结果抗血清的效价为 1∶30 0 0 0 ,交叉反应性除皮质酮为 0 .0 2 % ,其余均小于0 .0 1% ,批内差异为 7.8%～ 8.2 % ,批间差异为 9.9%～ 12 .1% ,平均回收率为 10 1.3% ,平行测定相关系数为 0 .9939(P <0 .0 1) ,最低可测定限为 10nmol/L ,正常参考值为 2 84 .7～ 16 9.2nmol/2 4h。原醛腺瘤组尿液 18-羟皮质醇明显高于原醛增生组、原发性高血压组和正常组 (P <0 .0 5 ) ,后三组之间无显著性差异 (P >0 .0 5 )。　结论本方法具有简便、灵敏、稳定等特点 ,可准确反应机体 2 4hObjective To develop a competitive avidin-biotin complex enzyme linked-immunoassay for 18-hydroxycortisol in urine. Methods The antisera of 18-hydroxycortisol were produced by immunizing the rabbit, and then the antisera was used to establish a competitive avidin-biotin enzyme linked-immunoassay (ABC-ELISA) for urinary 18-hydroxycortisol. Using the assay we measured the levels of 18-hydroxycortisol in urine in normal persons and patients with aldosterone-prodcuing adenoma (APA), idiopathic hyperaldosteronism (IHA) and essential hypertension. Results An antiserum with high specificity and high titer (1∶30?000) was obtained. Its cross-reactions with other steroids were less than 0.01%, except with corticosterone(0.02%). The mean rate of recovery of this assay was 101.3%; the intra- and inter-assay coefficients of variation were 7.0% ～ 8.2% and 9.9% ～ 12.1%, respectively. The detection limit of human urine was 20nmol/L. The normal value of 18-hydroxycortisol was 284.7 ～ 169.2nmol/24h. The level of 18-hydroxycortisol in patients with aldosterone-producing adenoma was significantly higher than that of normal and other groups of patients (P< 0.05). Unlike the APA group, the level of 18-hydroxycortisol in other groups was not different (P>0.05). Conclusion The ABC-ELISA for urinary 18-hydroxycortisol was convenient, reproducible and highly sensitive. It could be used as a highly sensitive and specific discriminator for the differential diagnosis of primary aldosteronism.

关 键 词：酶联免疫测定 18-羟皮质醇 原发性醛固酮增多症 诊断 治疗 

分 类 号：R586.24[医药卫生—内分泌] R446.122[医药卫生—内科学]