Devreese Katrien, Wijns Walter, Combes Isabelle, Van kerckhoven Soetkin, Hoylaerts Marc F
Coagulation Laboratory, Department of Clinical Chemistry, Microbiology and Immunology, Ghent University Hospital, Ghent, Belgium.
Thromb Haemost. 2007 Sep;98(3):600-13.
Coagulation tests and coagulation factor assays have been complemented recently with experimental tests to measure the total amount of thrombin formed. We have presently analyzed thrombin generation of healthy adult and paediatric plasma samples via a fluorogenic and a chromogenic method. The chromogenic method was performed on the fully automated Behring Coagulation System (BCS) and fluorogenic assays via Calibrated Automated Thrombography (CAT), after coagulation induction by various tissue factor (TF) concentrations. Sample distribution and variability were analyzed for the four main coagulation parameters, derived via computerized curve analysis in each method. Results for both methods were correlated. At the recommended TF concentration (300 pM), thrombin generation via BCS was less variable than via CAT (1-6 pM), but at comparable TF concentrations (1-6 pM), the CAT sensitivity was higher than that of BCS. Inhibition of intrinsic coagulation with the anti-factor VIII antibody BO2C11 revealed that the BCS detected extrinsic coagulation exclusively, at all TF concentrations tested. In contrast, at low TF concentrations (1 and 2.5 pM), via CAT, intrinsic coagulation pathway amplification was measured. At standardized TF concentrations (300 pM in BCS vs. 2.5 pM in CAT), different reference values between adults and children were found, for all parameters, except Tmax. In adult samples, the best correlation between both methods was observed for ETP(CAT) versus ETP(BCS) and for Peak height(CAT) versus Cmax(BCS), when thrombin generation was exclusively extrinsic (300 pM in BCS vs. 6 pM in CAT). In conclusion, differential thrombin generation characteristics in BCS and CAT are relevant for their clinical applicability.
凝血试验和凝血因子测定最近已通过实验测试得到补充，以测量形成的凝血酶总量。我们目前通过荧光法和显色法分析了健康成人和儿童血浆样本的凝血酶生成情况。显色法在全自动贝林凝血系统（BCS）上进行，荧光法通过校准自动血栓形成检测法（CAT）进行，在由不同组织因子（TF）浓度诱导凝血后进行。通过计算机曲线分析在每种方法中得出四个主要凝血参数，并对样本分布和变异性进行了分析。两种方法的结果相互关联。在推荐的TF浓度（300 pM）下，通过BCS产生的凝血酶变异性低于通过CAT产生的凝血酶（1 - 6 pM），但在相当的TF浓度（1 - 6 pM）下，CAT的灵敏度高于BCS。用抗因子VIII抗体BO2C11抑制内源性凝血表明，在所有测试的TF浓度下，BCS仅检测外源性凝血。相比之下，在低TF浓度（1和2.5 pM）下，通过CAT测量到了内源性凝血途径的放大。在标准化的TF浓度下（BCS中为300 pM，CAT中为2.5 pM），除Tmax外，所有参数在成人和儿童之间都发现了不同的参考值。在成人样本中，当凝血酶生成仅为外源性时（BCS中为300 pM，CAT中为6 pM），两种方法之间在ETP（CAT）与ETP（BCS）以及峰高（CAT）与Cmax（BCS）之间观察到了最佳相关性。总之，BCS和CAT中不同的凝血酶生成特征与其临床适用性相关。
