Sohn H E, Furukawa Y, Yumita S, Miura R, Yoshinaga K, Yamane R
Nihon Naibunpi Gakkai Zasshi. 1985 Sep 20;61(9):912-23. doi: 10.1507/endocrine1927.61.9_912.
Several problems in the measurement of plasma cyclic AMP (PcAMP) and nephrogenous cyclic AMP were studied using a YAMASA RIA Kit (YAMASA Shoyu, Choshi, Japan). In this assay method, cAMP in plasma is directly succinylated without prior deproteinization, and then it is bound to antibody in an imidazole buffer. So far as the blood samples were obtained with EDTA-4Na at least 5.0 mM in the final concentration, PcAMP was not reduced until 16 hours after the blood samples were drawn. Even without EDTA, the reductions in PcAMP were not detected within 1 hour after the blood samples were drawn. This assay method for PcAMP showed parallelism in the dilution curve. Recovery was almost complete. Intra- and interassay variations were low. When plasma was incubated at 37 degrees C for 24 hours, PcAMP became negligible. Furthermore, the values of PcAMP measured with this direct assay system almost agreed with those obtained after the purification by deproteinization and Dowex column chromatography through an anion-exchange resin. The normal values of PcAMP were 13.6 +/- 3.62 pmol/ml [mean +/- SD, n = 43]. Nephrogenous cAMP expressed as a function of GFR never did show any negative values in various clinical situations. From the data of basal levels and the oral calcium tolerance test, nephrogenous cAMP appeared to be more useful than total urinary cAMP in the diagnosis of parathyroid disorders, especially hyperparathyroidism.
使用日本铫子市的山正酱油株式会社生产的山正放射免疫分析试剂盒(YAMASA RIA Kit),对血浆环磷酸腺苷(PcAMP)和肾源性环磷酸腺苷的测量中的几个问题进行了研究。在这种检测方法中,血浆中的环磷酸腺苷无需预先去除蛋白就直接进行琥珀酰化,然后在咪唑缓冲液中与抗体结合。只要采集血样时最终浓度至少为5.0 mM的乙二胺四乙酸四钠(EDTA - 4Na),在采血后16小时内PcAMP都不会降低。即使没有EDTA,在采血后1小时内也未检测到PcAMP的降低。这种PcAMP的检测方法在稀释曲线上显示出平行性。回收率几乎是完全的。批内和批间变异都很低。当血浆在37℃孵育24小时时,PcAMP变得可以忽略不计。此外,用这种直接检测系统测得的PcAMP值几乎与通过去除蛋白和经阴离子交换树脂的Dowex柱色谱法纯化后得到的值一致。PcAMP的正常值为13.6±3.62 pmol/ml[平均值±标准差,n = 43]。在各种临床情况下,以肾小球滤过率(GFR)表示的肾源性环磷酸腺苷从未显示出任何负值。从基础水平数据和口服钙耐量试验来看,在甲状旁腺疾病尤其是甲状旁腺功能亢进的诊断中,肾源性环磷酸腺苷似乎比总尿环磷酸腺苷更有用。