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Calibrated Automated Thrombin Generation Measurement in Clotting PlasmaHemker H.C.a · Giesen P.a · Al Dieri R.a · Regnault V.b · de Smedt E.a · Wagenvoord R.a · Lecompte T.b · Béguin S.a,b
aSynapse BV, Cardiovascular Research Institute, University of Maastricht, Maastricht, The Netherlands; bINSERM ERIT-M 0323 et EA 3452, Faculté de Médecine, Université Henri Poincaré, Nancy, France Corresponding Author
H.C. Hemker, MD, PhD
CARIM, POB 616
NL–6200MD Maastricht (The Netherlands)
Tel. +31 43 3881 675, Fax +31 43 3884 570, HC.Hemker@Thrombin.com
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 ∼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.
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