Calibrated Automated Thrombogram (CAT Sample Clauses

Calibrated Automated Thrombogram (CAT. 1.5.5.2.1 The mechanism of the CAT Xxxxxx and colleagues have developed a method to measure thrombin generation which was reproducible and accurate. (Xxxxxx et al., 2003) The thrombin generation assay measures the ability of plasma to generate thrombin following in-vitro activation of coagulation with tissue factor. A thrombin generation curve is produced which reflects both the initiation and propagation phase of coagulation, as well as the anti-coagulant and fibrinolytic phases. (Xxxxxx et al., 2000) CAT measures thrombin generation in whole or platelet poor plasma. (Xxxxxxx et al., 2013) The CAT test is performed in triplicate on the sample and on the calibrator, using a microtitre plate. In the measurement xxxxx, tissue factor and phospholipid vesicles are added to initiate coagulation, then a combination of calcium chloride and fluorogenic substrate is added to the sample. A known amount of substrate converting activity is added to the plasma in the xxxxx and a low-affinity fluorogenic substrate for thrombin is added. Thrombin causes splitting of the substrate, thus, the thrombin activity in the clotting plasma is continuously monitored via the colour change. The developing fluorescence is recorded by a fluorometer and the fluorescence output values are converted into thrombin generation curves by an integrated software, comparing the fluorogenic activity to a constant known thrombin activity in a simultaneously run non-clotting sample. The calibration xxxxx contain the thrombin calibrator, which consists of thrombin bound to alpha2- macroglobulin, which cannot be split by plasma protease inhibitors, thus, these xxxxx do not clot. The calibration xxxxx have a standard known amount of substrate converting activity, thus, the fluorogenic substrate is converted at a constant rate by the added thrombin calibrator. (Xxxxxx et al., 2000; Xxxxxx et al., 2003; Xxxxxx, Xxxx, 2004) The parameters of the thrombin generation curve are described in terms of lag time, peak height, area under the curve, time to peak and start tail (figure 3).
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