Immunoluminometric assay for measurement of the C-terminal endothelin-1 precursor fragment in human plasma
Immunoluminometric assay for measurement of the C-terminal endothelin-1 precursor fragment in human plasma
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DOI：
10.1373/clinchem.2005.065581
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发表时间：
2006-06-01
影响因子：
9.3
通讯作者：
Bergmann, Andreas
中科院分区：
文献类型：
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作者：
Papassotiriou, Jana;Morgenthaler, Nils G.;Bergmann, Andreas
背景：内皮素-1(ET-1)是一种有效的血管收缩因子，由于其不稳定性及其与受体和血浆蛋白的结合，其测定很困难。我们报道了一种快速、可靠的间接定量ET-1释放的方法，该方法通过测量ET-1的C末端前体片段(CT-ProET-1)而无需提取步骤。方法：在健康人、充血性心力衰竭(CHF)患者和脓毒症患者的血浆样本中，我们用两种针对前ProET-1的168-212氨基酸的多克隆抗体的夹心免疫发光法测定CT-ProET-1。结果：CT-ProET-1浓度与BigET-1测定结果在3h内呈线性关系，检出限为0.4pmol/L，实验室间变异系数为10%。在EDTA、肝素和柠檬酸血浆中，在室温下至少4h，在4℃下6h，在EDTA血浆中，在-20℃下至少6个月，CT-ProET-1值在健康人中服从正态分布(平均值为44.3pmol/L；范围10.5-77.4pmol/L)，男女之间没有显著差异。CT-ProET-1与年龄的相关系数为0.0001(P<0.05)。慢性心力衰竭患者(中位数104pmol/L；范围50.8~315pmol/L)和败血症患者(中位数189pmol/L；范围34.6~855pmol/L)的CT-ProET1显著升高(P<；0.0001)。43份标本中CT-ProET-1与BigET-1的相关性为0.80(P<；0.0001)。结论：CT-ProET-1测定是间接评价危重病患者ET-1释放的一种快速、简便的方法。(C)2006年美国临床化学协会。
Background: Endothelin-1 (ET-1), a potent vasoconstrictor, is difficult to measure because of its instability and its binding to receptors and plasma proteins. We report a rapid, robust way to indirectly quantify ET-1 release by measuring the C-terminal ET-1 precursor fragment (CT-proET-1) without an extraction step.Methods: In plasma samples from healthy individuals, patients with congestive heart failure (CHF), and patients with sepsis, we measured CT-proET-1 with a sandwich immunoluminometric assay that uses 2 polyclonal antibodies to amino acids 168-212 of pre-proET-1. We also correlated CT-proET-1 concentrations with bigET-1 measurements.Results: The assay yielded results within 3 h and showed linear dilution with an analytical detection limit of 0.4 pmol/L and an interlaboratory CV < 10% for values > 10 pmol/L. Ex vivo CT-proET-1 was stable (< 10% loss of immunoreactivity) in EDTA-, heparin-, and citrate-plasma for at least 4 h at room temperature, 6 h at 4 degrees C, and in EDTA-plasma for at least 6 months at -20 degrees C. CT-proET-1 values followed a gaussian distribution in healthy individuals (mean, 44.3 pmol/L; range, 10.5-77.4 pmol/L) without significant differences between males and females. The correlation coefficient for CT-proET-1 vs age was 0.25 (P < 0.0001). CT-proET1 was significantly (P < 0.0001) increased in patients with CHF (median, 104 pmol/L; range, 50.8-315 pmol/L) and patients with sepsis (median, 189 pmol/L; range, 34.6-855 pmol/L). The correlation between CT-proET-1 and bigET-1 for 43 samples was 0.80 (P < 0.0001).Conclusions: CT-proET-1 measurement is a rapid and easy method for indirectly assessing the release of ET-1 in critically ill patients. (c) 2006 American Association for Clinical Chemistry.