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一种皮质酮直接放射免疫测定法的开发与应用。

The development and application of a direct radioimmunoassay for corticosterone.

作者信息

Al-Dujaili E A, Williams B C, Edwards C R

出版信息

Steroids. 1981 Feb;37(2):157-76. doi: 10.1016/s0039-128x(81)80015-3.

Abstract

A direct, simple and highly specific radioimmunoassay for corticosterone has been developed. The assay does not require preliminary solvent extraction of the sample or any chromatographic step. The assay utilises a highly specific antibody raised in rabbits against corticosterone-3-(0-carboxymethyl)-oxime-BSA immunogen and gamma-labeled corticosterone of high specific activity. An excellent correlation was obtained between results of the direct assay and those measured after paper chromatography (r = 0.99, P less than 0.001). The coefficients of variation for intra-assay and inter-assay determinations of samples from normal and high plasma pools were 4.6-6.2% and 6.4-8.2% respectively. The minimum limit of detection was 5 pg/assay tube (0.1 ng/ml). The assay has been applied to assess plasma corticosterone levels in various physiopathological and pathophysiological studies. It is extremely practical to the extent that a single technician can assay up to 1000 samples in a working week. Finally, the direct assay has been validated and employed for in vitro adrenal superfusion studies using either rat or human adrenal cells. The large numbers of samples produced by these studies would have exceeded the capacity of earlier radioimmunoassays requiring initial extraction and chromatography.

摘要

已开发出一种用于皮质酮的直接、简单且高度特异的放射免疫测定法。该测定法无需对样品进行初步溶剂萃取或任何色谱步骤。该测定法利用在兔体内产生的针对皮质酮 - 3 -(邻 - 羧甲基)- 肟 - BSA免疫原的高度特异性抗体以及高比活性的γ标记皮质酮。直接测定结果与纸色谱后测定结果之间获得了极好的相关性(r = 0.99,P小于0.001)。来自正常和高血浆池的样品的批内和批间测定的变异系数分别为4.6 - 6.2%和6.4 - 8.2%。最低检测限为5 pg/测定管(0.1 ng/ml)。该测定法已应用于各种生理病理和病理生理研究中评估血浆皮质酮水平。其极为实用,以至于一名技术人员在一个工作周内最多可测定1000个样品。最后,该直接测定法已通过验证,并用于使用大鼠或人肾上腺细胞的体外肾上腺灌流研究。这些研究产生的大量样品将超出早期需要初始萃取和色谱的放射免疫测定法的能力。

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