Feinglos M N, Drezner M K, Lebovitz H E
J Clin Endocrinol Metab. 1978 May;46(5):824-9. doi: 10.1210/jcem-46-5-824.
Currently used methods for plasma cAMP measurements are either tedious (chromatographic preparation of sample) or potentially inaccurate (direct assay of plasma samples). A rapid, simple, and accurate competitive binding assay for plasma cAMP, which does not require chromatographic preparation of the sample, has been developed. This procedure prevents destruction of plasma cAMP by utilizing both theophylline and EDTA in the collection of the blood sample. Human plasma contains variable amounts of cAMP-binding activity which interfere with the measurement of cAMP by the standard competitive binding assay. Our assay procedure removes this binding activity by precipitation of plasma proteins with perchloric acid. The normal fasting value (+/- SD) of plasma cAMP using this technique is 17.6 +/- 4.3 pmol/ml, which is identical to values obtained by methods utilizing chromatographic purification of samples (18.3 +/- 3.0). The fasting plasma cAMP of patients with hyperparathyroidism is normal (16.2 +/- 3.4), but patients with maturity-onset diabetes mellitus have fasting values significantly below normal (12.3 +/- 2.4).
目前用于测量血浆环磷酸腺苷(cAMP)的方法要么繁琐(样品的色谱制备),要么可能不准确(血浆样品的直接测定)。现已开发出一种快速、简单且准确的血浆cAMP竞争性结合测定法,该方法无需对样品进行色谱制备。此程序通过在采集血样时同时使用茶碱和乙二胺四乙酸(EDTA)来防止血浆cAMP被破坏。人血浆中含有可变数量的cAMP结合活性,这会干扰通过标准竞争性结合测定法对cAMP的测量。我们的测定程序通过用高氯酸沉淀血浆蛋白来去除这种结合活性。使用该技术测得的血浆cAMP正常空腹值(±标准差)为17.6±4.3皮摩尔/毫升,这与通过样品色谱纯化方法获得的值(18.3±3.0)相同。甲状旁腺功能亢进患者的空腹血浆cAMP正常(16.2±3.4),但成年型糖尿病患者的空腹值明显低于正常水平(12.3±2.4)。