Hemker H Coenraad, Giesen Peter, Al Dieri Raed, Regnault Véronique, de Smedt Eric, Wagenvoord Rob, Lecompte Thomas, Béguin Suzette
Synapse BV, Cardiovascular Research Institute, University of Maastricht, Maastricht, The Netherlands.
Pathophysiol Haemost Thromb. 2003;33(1):4-15. doi: 10.1159/000071636.
Calibrated automated thrombography displays the concentration of thrombin in clotting plasma with or without platelets (platelet-rich plasma/platelet-poor plasma, PRP/PPP) in up to 48 samples by monitoring the splitting of a fluorogenic substrate and comparing it to a constant known thrombin activity in a parallel, non-clotting sample. Thus, the non-linearity of the reaction rate with thrombin concentration is compensated for, and adding an excess of substrate can be avoided. Standard conditions were established at which acceptable experimental variation accompanies sensitivity to pathological changes. The coefficients of variation of the surface under the curve (endogenous thrombin potential) are: within experiment approximately 3%; intra-individual: <5% in PPP, <8% in PRP; interindividual 15% in PPP and 19% in PRP. In PPP, calibrated automated thrombography shows all clotting factor deficiencies (except factor XIII) and the effect of all anticoagulants [AVK, heparin(-likes), direct inhibitors]. In PRP, it is diminished in von Willebrand's disease, but it also shows the effect of platelet inhibitors (e.g. aspirin and abciximab). Addition of activated protein C (APC) or thrombomodulin inhibits thrombin generation and reflects disorders of the APC system (congenital and acquired resistance, deficiencies and lupus antibodies) independent of concomitant inhibition of the procoagulant pathway as for example by anticoagulants.
校准自动血栓形成检测法通过监测荧光底物的裂解，并将其与平行非凝血样本中已知的恒定凝血酶活性进行比较，来显示有或无血小板（富血小板血浆/贫血小板血浆，PRP/PPP）的凝血血浆中凝血酶的浓度，最多可检测48个样本。因此，反应速率与凝血酶浓度的非线性关系得到补偿，且可避免添加过量底物。建立了标准条件，在此条件下，对病理变化的敏感性伴随着可接受的实验变异。曲线下面积（内源性凝血酶潜力）的变异系数为：实验内约3%；个体内：PPP中<5%，PRP中<8%；个体间PPP中为15%，PRP中为19%。在PPP中，校准自动血栓形成检测法可显示所有凝血因子缺乏（因子 XIII 除外）以及所有抗凝剂[维生素 K 拮抗剂（AVK）、肝素（类肝素）、直接抑制剂]的作用。在PRP中，血管性血友病时其作用减弱，但也可显示血小板抑制剂（如阿司匹林和阿昔单抗）的作用。添加活化蛋白 C（APC）或血栓调节蛋白可抑制凝血酶生成，并反映 APC 系统的紊乱（先天性和获得性抵抗、缺乏及狼疮抗体），且与例如抗凝剂对促凝途径的伴随抑制无关。
