Beck Karl-Heinz
Department of Transfusion Medicine and Hemostaseology, University Hospital of Freiburg, Schönbergweg 16, 79283 Bollschweil/Freiburg, Breisgau, Germany.
Transfus Apher Sci. 2002 Dec;27(3):247-53. doi: 10.1016/s1473-0502(02)00072-1.
The PFA-100 examines platelet function by measuring closure time (CT) when whole blood is passing through a capillary and a filter membrane coated with a collagen/platelet agonist. Termination of the CT is achieved when adhered and aggregated platelets of the passing blood flow occlude the membrane. Two PFA-100 test cartridges are used to measure the CT of citrated whole blood; either the collagen/ADP (PCA) or the collagen/ epinephrine (PCE). We investigated the applicability of the PFA-100 test system for quality control of platelet concentrates in comparison with platelet aggregometry.
For platelet aggregation, the combination of ADP (100 microM/l) and collagen (20 microg/ml) were used as agonists (ADP/COL). In our test system, 25 microl epinephrine (10(-3) M) were added to the PCA cartridge (CT: PCAE) and 25 microl ADP (100 microM/l) to the PCE cartridge (CT: PCEA), respectively. Red blood cells from a blood group 0 donor were adjusted to a hematocrit of 43% using platelet rich plasma (8 x 10(5)/microl) of the respective platelet concentrate. Fourteen irradiated and non-irradiated platelet concentrates were examined on days 0, 3 and 5 after platelet preparation. Swirling without a scoring system and mean platelet volume (MPV) were also tested. Statistical analyses were performed by Pearson's range coefficient, the Mann and Whitney test, and the Wilcoxon test.
Swirling was seen in all platelet concentrates. Mean platelet volume was normal during the whole observation period like the PCAE in non-irradiated products. Only the PCAE showed finite CTs during the whole storage time. Consequently, further testing was performed exclusively with the PCAE. Weak but significant correlations could be found between the PCAE and ADP/COL (r = -0.53 for non-irradiated platelet products, r = -0.62 for irradiated platelet products, p < or = 0.0001). Platelet function significantly decreased to around 60% of the initial value in the PCAE on day 5 of storage, ADP/COL decreased to 35% of maximum amplitude. A further decrease of approximately 10% was observed in the irradiated products, a fact which was significant in the PCAE. Only in the irradiated platelet products did MPV show a weak but significant correlation with the PCAE (r = 0.45, p = 0.002).
Functional features of platelets from platelet concentrates characterized by ADP/COL induced aggregation can also be measured by the PFA-100. Yet, it has a higher sensitivity to platelet lesions induced by gamma-radiation. Further clinical studies will be necessary to determine whether PFA-measurements meet the quality criteria of MPV or swirling, or is superior in predicting in vivo viability. The relatively high cost of the test cartridge remains an obstacle for routine use.
PFA-100通过测量全血通过涂有胶原蛋白/血小板激动剂的毛细血管和滤膜时的封闭时间(CT)来检测血小板功能。当流经的血流中黏附并聚集的血小板阻塞滤膜时,CT结束。使用两个PFA-100测试盒来测量枸橼酸化全血的CT;即胶原蛋白/ADP(PCA)或胶原蛋白/肾上腺素(PCE)。我们将PFA-100测试系统与血小板聚集测定法进行比较,研究其在血小板浓缩物质量控制中的适用性。
对于血小板聚集,使用ADP(100微摩尔/升)和胶原蛋白(20微克/毫升)的组合作为激动剂(ADP/COL)。在我们的测试系统中,分别向PCA测试盒(CT:PCAE)中加入25微升肾上腺素(10⁻³摩尔/升),向PCE测试盒(CT:PCEA)中加入25微升ADP(100微摩尔/升)。使用相应血小板浓缩物的富血小板血浆(8×¹⁰⁵/微升)将0型供血者的红细胞调整至血细胞比容为43%。在血小板制备后的第0天、第3天和第5天,对14份辐照和未辐照的血小板浓缩物进行检测。还测试了无评分系统的漩涡现象和平均血小板体积(MPV)。采用Pearson极差系数、Mann和Whitney检验以及Wilcoxon检验进行统计分析。
在所有血小板浓缩物中均观察到漩涡现象。在整个观察期内,未辐照产品中的平均血小板体积与PCAE一样正常。仅PCAE在整个储存期内显示出有限的CT值。因此,仅对PCAE进行了进一步检测。在PCAE与ADP/COL之间可发现弱但显著的相关性(未辐照血小板产品中r = -0.53,辐照血小板产品中r = -0.62,p≤0.0001)。在储存第5天时,PCAE中血小板功能显著下降至初始值的约60%,ADP/COL下降至最大振幅的35%。在辐照产品中观察到进一步下降约10%,这在PCAE中具有显著性。仅在辐照血小板产品中,MPV与PCAE显示出弱但显著的相关性(r = 0.45,p = 0.002)。
以ADP/COL诱导聚集为特征的血小板浓缩物中血小板的功能特性也可通过PFA-100进行测量。然而,它对γ射线诱导的血小板损伤具有更高的敏感性。需要进一步的临床研究来确定PFA测量是否符合MPV或漩涡现象的质量标准,或者在预测体内活力方面是否更优越。测试盒相对较高的成本仍然是常规使用的障碍。