Nilsson Torbjörn K, Boman Kurt, Jansson Jan-Håkan, Thøgersen Anna Margrethe, Berggren Mona, Broberg Annette, Granlund Anna
Department of Clinical Chemistry, Orebro University Hospital, Orebro, Sweden.
Thromb Res. 2005;116(3):249-54. doi: 10.1016/j.thromres.2004.12.005. Epub 2005 Jan 12.
The aim was to define the most suitable specimen collection tubes for measurements of soluble Thrombomodulin (sTM), von Willebrand factor (vWF), and tPA/PAI-1 complex concentrations, and in particular whether the strongly acidic citrate additive in Stabilyte plasma would give significantly improved long-term stability of any of these analytes. We measured these analytes in paired specimens from 34 subjects, sampled 8-11 years before analysis, in serum, EDTA plasma, citrated plasma, and acidified citrated plasma (Stabilyte). Results were evaluated by regression analysis and Bland-Altman plots. All associations were linear across a wide assay range. Soluble TM was found to be highly unstable in serum as well as in EDTA plasma and to some extent even in ordinary citrate plasma: acidified citrate plasma is necessary to preserve sTM immunoreactivity in long-term storage. For hsCRP the slopes were not significantly different from that predicted by the dilution effect (0.83-0.86) of the citrate additive and there was no appreciable intercept. vWF values were comparable in citrate and acidified citrate plasma but serum and EDTA plasma samples yielded lower than expected results. For tPA/PAI-1 complex, Stabilyte tubes gave systematically lower results than the other tubes, with serum and EDTA plasma scoring the highest values, suggesting that in vitro increase in complex levels takes places upon blood collection and/or storage. We conclude that Stabilyte plasma is the specimen collection tube of choice for biobank projects aiming to measure fibrinolytic factors as well as several other analytes in the clotting system, such as soluble thrombomodulin and von Willebrand factor, in addition to the inflammatory marker hs-CRP. Indeed, using acidified Stabilyte plasma as the single medium would substantially simplify sampling for many epidemiological studies.
目的是确定用于测量可溶性血栓调节蛋白(sTM)、血管性血友病因子(vWF)和tPA/PAI-1复合物浓度的最合适的标本采集管,特别是Stabilyte血浆中强酸性柠檬酸盐添加剂是否会显著提高这些分析物中任何一种的长期稳定性。我们在分析前8 - 11年采集的34名受试者的配对标本中测量了这些分析物,标本类型包括血清、乙二胺四乙酸(EDTA)血浆、枸橼酸盐血浆和酸化枸橼酸盐血浆(Stabilyte)。通过回归分析和Bland-Altman图对结果进行评估。在广泛的检测范围内,所有相关性均呈线性。发现可溶性TM在血清以及EDTA血浆中高度不稳定,甚至在普通枸橼酸盐血浆中在一定程度上也不稳定:酸化枸橼酸盐血浆对于长期保存sTM免疫反应性是必要的。对于高敏C反应蛋白(hsCRP),其斜率与柠檬酸盐添加剂的稀释效应(0.83 - 0.86)预测的斜率无显著差异,且无明显截距。vWF值在枸橼酸盐血浆和酸化枸橼酸盐血浆中相当,但血清和EDTA血浆样本的结果低于预期。对于tPA/PAI-1复合物,Stabilyte管给出的结果系统地低于其他管,血清和EDTA血浆的值最高,这表明在血液采集和/或储存时复合物水平在体外会升高。我们得出结论,对于旨在测量凝血系统中的纤溶因子以及其他几种分析物(如可溶性血栓调节蛋白和血管性血友病因子)以及炎症标志物hs-CRP的生物样本库项目,Stabilyte血浆是标本采集管的首选。事实上,使用酸化的Stabilyte血浆作为单一介质将大大简化许多流行病学研究的采样。