血液和血浆样本中体外补体激活的可能机制：富山/乙二胺四乙酸可控制体外补体激活。
Possible mechanism for in vitro complement activation in blood and plasma samples: futhan/EDTA controls in vitro complement activation.
作者信息
Pfeifer P H, Kawahara M S, Hugli T E
机构信息
The Futhan/EDTA tubes are produced by Terumo Europe and can be purchased from PerSeptive Biosystems, 500 Old Connecticut Path, Framingham, MA 01701, USA.
出版信息
Clin Chem. 1999 Aug;45(8 Pt 1):1190-9.
BACKGROUND
Ongoing in vitro complement (C) activation in citrate or EDTA plasma has prevented an accurate analysis of C-activation products generated in vivo. The aim of this study was to characterize handling and storage conditions required to prevent in vitro C activation in blood and plasma samples collected with Futhan/EDTA.
METHODS
Biotrak(TM) RIAs were used to quantitatively measure C3a and C4a in blood and/or plasma samples from healthy individuals (controls) and from liver transplant patients. Blood samples were routinely drawn into either EDTA (1 g/L) tubes or into tubes containing both EDTA (1 g/L) and Futhan (0.1 g/L) and immediately centrifuged at 2000g for 15 min at 4 degrees C.
RESULTS
In controls, C4a, but not C3a, in fresh samples (time 0) was higher in EDTA plasma than in Futhan/EDTA plasma (n = 20; P = 0.002). Futhan/EDTA prevented C3a and C4a generation in blood and plasma samples held at room temperature (22-23 degrees C) for 1 h and in plasma held for 24 h at 4 degrees C or -70 degrees C. The mean C3a concentration (1.76 mg/L; n = 19) at time 0 in EDTA plasma samples from liver transplant patients was significantly higher than for controls (0.34 mg/L; n = 11). In these patients, the mean C3a in EDTA samples increased to 13.8 mg/L after 60 min at room temperature, but there was no change in the C3a concentration of an EDTA plasma from a control. In the patients, C3a concentrations were lower in Futhan/EDTA plasma than in EDTA at time 0 and after 60 min at room temperature (1.40 and 2.02 mg/L, respectively). The mean patient C4a was 4.02 mg/L in EDTA plasma at time 0 vs 0.24 mg/L for controls; it increased to 16.9 mg/L after 60 min at room temperature compared with 0.76 mg/L for controls. The mean patient C4a was 0.83 mg/L in Futhan/EDTA plasma at time 0 vs 0.1 mg/L for controls. Neither patient nor control C4a concentrations increased vs time in Futhan/EDTA.
CONCLUSION
The combination of Futhan (0.1 g/L) and EDTA (1 g/L) eliminates in vitro C activation.
背景
柠檬酸盐或乙二胺四乙酸（EDTA）血浆中持续的体外补体（C）激活阻碍了对体内产生的补体激活产物的准确分析。本研究的目的是确定防止用氟烷/EDTA采集的血液和血浆样本发生体外补体激活所需的处理和储存条件。
方法
使用Biotrak™放射免疫分析法（RIA）定量测量健康个体（对照组）和肝移植患者的血液和/或血浆样本中的C3a和C4a。血液样本常规采集到含有EDTA（1 g/L）的试管中，或采集到同时含有EDTA（1 g/L）和氟烷（0.1 g/L）的试管中，并立即在4℃下以2000g离心15分钟。
结果
在对照组中，新鲜样本（时间0）中，EDTA血浆中的C4a而非C3a高于氟烷/EDTA血浆（n = 20；P = 0.002）。氟烷/EDTA可防止血液和血浆样本在室温（22 - 23℃）下保存1小时以及血浆在4℃或 - 70℃下保存24小时期间产生C3a和C4a。肝移植患者的EDTA血浆样本在时间0时的平均C3a浓度（1.76 mg/L；n = 19）显著高于对照组（0.34 mg/L；n = 11）。在这些患者中，EDTA样本在室温下放置60分钟后，平均C3a浓度增加到13.8 mg/L，但对照组的EDTA血浆中C3a浓度没有变化。在患者中，氟烷/EDTA血浆在时间0时以及在室温下放置60分钟后的C3a浓度低于EDTA血浆（分别为1.40和2.02 mg/L）。患者的EDTA血浆在时间0时平均C4a为4.02 mg/L，而对照组为0.24 mg/L；与对照组的0.76 mg/L相比，在室温下放置60分钟后增加到16.9 mg/L。患者的氟烷/EDTA血浆在时间0时平均C4a为0.83 mg/L，而对照组为0.1 mg/L。在氟烷/EDTA中，患者和对照组的C4a浓度均未随时间增加。
结论
氟烷（0.1 g/L）和EDTA（1 g/L）的组合可消除体外补体激活。
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