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首页> 外文期刊>Journal of immunoassay >One step enzyme linked immunosorbent assay for direct estimation of serum cortisol.
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One step enzyme linked immunosorbent assay for direct estimation of serum cortisol.

机译：一步酶联免疫吸附法直接估算血清皮质醇。

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摘要

One step competitive enzyme linked immunosorbent assay (ELISA) for direct estimation of cortisol in human serum is described. Cortisol-3-O-carboxymethyl-oxime-bovine serum albumin (cortisol-3-O-CMO-BSA) was used as an immunogen and cortisol-21-hemisuccinate-horse radish peroxidase (cortisol-21-HS-HRP) was used as a tracer. To the cortisol antibody coated microtiter wells, standards or serum samples (25 microl) along with cortisol-HRP conjugate (100 microl) were incubated for 2 hours at 37 degrees C. Bound enzyme activity was measured by, using TMB/H2O2 as a substrate. In this new strategy, chilled acetone stripped pooled human serum and sodium salicylate were used for preparing the standards and blocking the cortisol binding globulin (CBG), respectively. The sensitivity of the assay was .28 microg/100ml. The intraassay and interassay coefficient of variations (CVs) were ranged from 1.3% to 9.3% and 6.8% to 12.3 %, respectively. The analytical recoveries were 94% to 101.5%. The serum cortisol values, obtained by this method were correlated well with those, obtained by radioimmunoassay; r=0.95 (n=52).
机译：描述了用于直接估算人血清中皮质醇的一步竞争性酶联免疫吸附测定（ELISA）。使用Cortisol-3-O-羧甲基肟肟-牛血清白蛋白（cortisol-3-O-CMO-BSA）作为免疫原，并使用cortisol-21-半琥珀酸酯-马萝卜过氧化物酶（cortisol-21-HS-HRP）作为示踪剂。将标准或血清样品（25微升）与皮质醇-HRP共轭物（100微升）一起在37摄氏度下温育2小时。在皮质醇抗体包被的微量滴定孔中。以TMB / H2O2为底物测量结合的酶活性。在这种新策略中，将冷冻的丙酮剥离的合并的人血清和水杨酸钠用于制备标准液并分别阻断皮质醇结合球蛋白（CBG）。该测定法的灵敏度为0.28微克/ 100毫升。批内和批间变异系数（CVs）分别为1.3％至9.3％和6.8％至12.3％。分析回收率为94％至101.5％。通过这种方法获得的血清皮质醇值与通过放射免疫分析获得的血清皮质醇值具有良好的相关性; r ＝ 0.95（n ＝ 52）。

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