Watanabe F, Ryota T, Yamanaga H, Kobayashi Y, Ogihara T, Saeki S, Kumahara Y
Nihon Naibunpi Gakkai Zasshi. 1985 Oct 20;61(10):1157-66. doi: 10.1507/endocrine1927.61.10_1157.
An enzyme immunoassay for serum 18-hydroxycorticosterone was established using alkaline phosphatase as a label. The antiserum for 18-hydroxycorticosterone was produced by immunization of rabbits with 18-hydroxycorticosterone 3-(o-carboxymethyl) oxime conjugated to bovine serum albumin. Sephadex LH-20 column chromatography was used to separate 18-hydroxycorticosterone from other steroids in serum samples. The minimal detectable amount of 18-hydroxycorticosterone was 50 pg/tube, and the measurable range was from 5 to 1000 ng/dl when a 1.0 ml serum sample was used. Intra- and inter-assay coefficients of variance were 5.0% (n = 6), and 5.8% (n = 6), respectively. In normal controls the serum 18-hydroxycorticosterone level was 4.8 approximately 34.0 ng/dl (mean +/- S.D. = 17.1 +/- 9.0 ng/dl) on an unrestricted diet. Seven out of 8 patients with aldosterone-producing adenoma had above-normal serum 18-hydroxycorticosterone levels. Serum 18-hydroxycorticosterone increased and decreased significantly following ACTH and dexamethasone administration, respectively. In essential hypertensive patients, serum 18-hydroxycorticosterone was high during a low-sodium diet and was suppressed remarkably by captopril. These observations support the previous reports that adrenal 18-hydroxycorticosterone synthesis is dependent on both ACTH and the renin-angiotensin system. The present method is sufficiently sensitive and producible, avoids the use of radioisotopes and is quite satisfactory for clinical use.
建立了一种以碱性磷酸酶为标记物的血清18-羟皮质酮酶免疫测定法。用与牛血清白蛋白偶联的18-羟皮质酮3-(邻羧甲基)肟免疫家兔制备18-羟皮质酮抗血清。采用Sephadex LH-20柱色谱法从血清样本中的其他类固醇中分离18-羟皮质酮。18-羟皮质酮的最小可检测量为50 pg/管,当使用1.0 ml血清样本时,可测量范围为5至1000 ng/dl。批内和批间变异系数分别为5.0%(n = 6)和5.8%(n = 6)。在正常对照组中,自由饮食时血清18-羟皮质酮水平为4.8至约34.0 ng/dl(均值±标准差 = 17.1±9.0 ng/dl)。8例醛固酮分泌性腺瘤患者中有7例血清18-羟皮质酮水平高于正常。分别给予促肾上腺皮质激素(ACTH)和地塞米松后,血清18-羟皮质酮显著升高和降低。在原发性高血压患者中,低钠饮食时血清18-羟皮质酮升高,卡托普利可显著抑制其水平。这些观察结果支持了先前的报道,即肾上腺18-羟皮质酮的合成依赖于ACTH和肾素-血管紧张素系统。本方法灵敏度足够且可重复,避免了使用放射性同位素,临床应用效果相当满意。