Kohen F, Pazzagli M, Kim J B, Lindner H R
Steroids. 1980 Oct;36(4):421-37. doi: 10.1016/0039-128x(80)90030-6.
An immunoassay procedure for the determination of cortisol in human plasma is described, which utilizes chemiluminescence as the end point. A cortisol-isoluminol conjugate serves as the chemiluminescent marker. The light emission by this conjugate upon oxidation is delayed by prior incubation with anti-cortisol IgG, but not by unrelated gamma-globulin. This delayed light emission was inhibited by cortisol in a dose-dependent manner, with a linear range of 20-1000 pg steroid/assay tube. A competitive protein binding assay based on this procedure was applied to methylene chloride extracts of cortisol from normal and pathological human plasma (2-40 micrograms/100 ml). Cortisol values obtained by this procedure agreed well with those obtained by radioimmunoassay, using the same antiserum with tritiated cortisol as the label (r = 0.98). The chemiluminescence immunoassay is comparable to radioimmunoassay with regard to sensitivity, specificity, precision and accuracy. The advantage of the new assay procedure is that it obviates the need for counting radioactivity and for separation of bound and free ligand.
本文描述了一种用于测定人血浆中皮质醇的免疫分析方法，该方法以化学发光作为终点检测手段。皮质醇 - 异鲁米诺偶联物用作化学发光标记物。该偶联物在氧化时发出的光，在与抗皮质醇IgG预先孵育后会延迟，但与无关的γ球蛋白孵育则不会延迟。这种延迟发光会被皮质醇以剂量依赖的方式抑制，线性范围为20 - 1000 pg类固醇/测定管。基于此方法的竞争性蛋白结合分析应用于正常和病理性人血浆中皮质醇的二氯甲烷提取物（2 - 40微克/100毫升）。通过该方法获得的皮质醇值与使用相同抗血清、以氚化皮质醇作为标记物的放射免疫分析结果非常吻合（r = 0.98）。在灵敏度、特异性、精密度和准确性方面，化学发光免疫分析与放射免疫分析相当。新分析方法的优点是无需进行放射性计数以及分离结合型和游离型配体。
