Evaluation of enzyme immunoassay and radioimmunoassay methods for the measurement of plasma oxytocin.
Evaluation of enzyme immunoassay and radioimmunoassay methods for the measurement of plasma oxytocin.
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DOI：
10.1097/psy.0b013e31821df0c2
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发表时间：
2011-06
影响因子：
3.3
通讯作者：
Mendez AJ
中科院分区：
文献类型：
--
作者：
Szeto A;McCabe PM;Nation DA;Tabak BA;Rossetti MA;McCullough ME;Schneiderman N;Mendez AJ
人们越来越关注测量外周催产素水平，以更好地了解这种肽在哺乳动物行为，生理和疾病中的作用。本研究的目的是比较使用市售酶免疫测定法（EIA）和放射免疫测定法（RIA）测量血浆催产素的方法，以评估样品提取的必要性，并评估测定的免疫特异性。在提取和未提取的人血浆样品（n = 39）中测量催产素。采用高效液相色谱法和凝胶过滤色谱法分离催产素及其降解产物，然后用EIA或RIA法测定催产素的免疫反应峰。在没有提取的情况下，EIA测量的血浆比提取的血浆高100倍以上，提取和未提取血浆中催产素水平之间的相关性很小（斯皮尔曼rho =-0.10，p = 0.54）。使用RIA，大多数样品（> 90%）在提取或未提取的情况下低于检测水平。在样品提取物的色谱分离后，发现除了催产素之外还存在多种免疫反应性产物，这对测定的特异性产生了怀疑。催产素水平的变化在社会和行为挑战研究中有报道。本研究表明，样品提取是获得有效测定结果所必需的。催产素降解产物的变化可能有助于先前观察到的循环催产素水平对行为和社会挑战的反应。迫切需要有效和可靠的方法来测量生物样品中的催产素。
There is increased interest in measuring peripheral oxytocin levels to better understand the role of this peptide in mammalian behavior, physiology, and disease. The purpose of this study was to compare methods for plasma oxytocin measurement using a commercially available enzyme immunoassay (EIA) and radioimmunoassay (RIA), to evaluate the need for sample extraction, and to assess the immunospecificity of the assays. Oxytocin was measured in extracted and unextracted human plasma samples (n = 39). Oxytocin and its degradation products were separated by high performance liquid chromatography and gel filtration chromatography and then assayed by EIA or RIA to identify oxytocin immunoreactive peaks. Without extraction, plasma measured by EIA was more than 100-fold higher than in extracted plasma, and the correlation between oxytocin levels in extracted and unextracted plasma was minimal (Spearman’s rho = −0.10, p = 0.54). Using the RIA, most samples (> 90%) were below the level of detection with or without extraction. Following chromatographic fractionation of sample extracts, multiple immunoreactive products were found to be present in addition to oxytocin, which casts doubts on the specificity of the assays. Changes in oxytocin levels have been reported in social and behavioral challenge studies. This study indicates that sample extraction is necessary to obtain valid assay results. Changes in oxytocin degradation products are likely to contribute to the previously observed responses in circulating oxytocin levels to behavioral and social challenge. There is a critical need for validated and reliable methods to measure oxytocin in biological samples.