De Jongh R, Vranken J, Vundelinckx G, Bosmans E, Maes M, Heylen R
Department of Anaesthesia, Intensive Care Medicine and Emergency Medicine, Ziekenhuis Oost-Limburg, Genk, Belgium.
Cytokine. 1997 Sep;9(9):696-701. doi: 10.1006/cyto.1997.0217.
Levels of plasma cytokines, their receptors or immune-related peptides, are used in experimental and clinical medicine. However, no standards are available regarding the use of anticoagulants or blood processing including the technique of blood collection or time delay between blood sampling and centrifugation. Blood was collected from 10 patients in order to assay interleukin 6, soluble interleukin 6 receptor, soluble interleukin 2 receptor and soluble transferrin receptor by enzyme-linked immunosorbent assay (ELISA). At the time of sampling, five different collecting tubes were used: allowing coagulation, in the presence of EDTA, with EDTA being administered to the serum after its separation, in citrated and in heparinized blood. Blood was centrifuged 10 min after collection and separated serum or plasma was immediately frozen at -20 degrees C. To study the effect of time delay between blood sampling and processing, blood from 11 other patients was sampled, allowing coagulation; or in EDTA tubes and stored at 4 degrees C. The blood was centrifuged 10 min, 8 h and 24 h later, and the separated serum or EDTA plasma was stored at -20 degrees C until thawed for protein determination. Plasma interleukin 6 and soluble interleukin 6 receptor concentrations did not depend on the type of anticoagulant used or the time delay between sampling and processing. Soluble interleukin 2 receptor concentrations were not influenced by time delay before centrifugation, but concentrations were increased 10-fold in EDTA plasma exclusively when EDTA had contact with blood cells. Soluble transferrin receptor concentrations rose progressively with storage time before centrifugation. Contact of EDTA to whole blood or serum resulted in the same increase of soluble transferrin receptor concentration. The results suggest that standardization of the use of anticoagulant and time delay before centrifugation and serum or plasma separation is necessary for soluble interleukin 2 receptor and soluble transferrin receptor.
血浆细胞因子、其受体或免疫相关肽的水平被应用于实验医学和临床医学。然而，在抗凝剂的使用或血液处理方面，包括采血技术或采血与离心之间的时间间隔，尚无相关标准。为了通过酶联免疫吸附测定（ELISA）法检测白细胞介素6、可溶性白细胞介素6受体、可溶性白细胞介素2受体和可溶性转铁蛋白受体，从10名患者采集了血液。采样时，使用了五种不同的采血管：允许凝血的、存在乙二胺四乙酸（EDTA）的、EDTA在血清分离后加入的、枸橼酸盐抗凝的和肝素抗凝的血液。采血后10分钟进行离心，分离出的血清或血浆立即在-20℃冷冻。为了研究采血与处理之间时间间隔的影响，从另外11名患者采集血液，允许凝血；或采集到EDTA管中并在4℃保存。血液分别在10分钟、8小时和24小时后进行离心，分离出的血清或EDTA血浆在-20℃保存，直至解冻用于蛋白质测定。血浆白细胞介素6和可溶性白细胞介素6受体浓度不取决于所用抗凝剂的类型或采样与处理之间的时间间隔。可溶性白细胞介素2受体浓度不受离心前时间间隔的影响，但仅当EDTA与血细胞接触时，EDTA血浆中的浓度会增加10倍。可溶性转铁蛋白受体浓度在离心前随保存时间逐渐升高。EDTA与全血或血清接触会导致可溶性转铁蛋白受体浓度出现相同程度的升高。结果表明，对于可溶性白细胞介素2受体和可溶性转铁蛋白受体，抗凝剂的使用以及离心和血清或血浆分离前的时间间隔标准化是必要的。
