title: Calibrated automated thrombin generation measurement in clotting plasma.
abstract: 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.
authors.affiliation: Synapse BV, Cardiovascular Research Institute, University of Maastricht, Maastricht, The Netherlands. HC.Hemker@Thrombin.com
authors.forename: H Coenraad
authors.initials: HC
authors.lastname: Hemker
authors.forename: Peter
authors.initials: P
authors.lastname: Giesen
authors.forename: Raed
authors.initials: R
authors.lastname: Al Dieri
authors.forename: Véronique
authors.initials: V
authors.lastname: Regnault
authors.forename: Eric
authors.initials: E
authors.lastname: de Smedt
authors.forename: Rob
authors.initials: R
authors.lastname: Wagenvoord
authors.forename: Thomas
authors.initials: T
authors.lastname: Lecompte
authors.forename: Suzette
authors.initials: S
authors.lastname: Béguin
mesh_terms: D000925:Anticoagulants / Q000494:pharmacology; D019540:Area Under Curve; D001331:Automation; D001778:Blood Coagulation Disorders / Q000097:blood / Q000175:diagnosis; D001780:Blood Coagulation Tests / Q000295:instrumentation / Q000379:methods*; D002138:Calibration; D003374:Coumarins / Q000032:analysis; D005340:Fibrinogen / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000032:analysis; D005470:Fluorometry / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D009842:Oligopeptides / Q000032:analysis; D010949:Plasma; D011486:Protein C / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D012016:Reference Values; D012984:Software; D013917:Thrombin / Q000032:analysis / Q000096:biosynthesis*; D018180:Thrombomodulin / Q000737:chemistry; D013925:Thromboplastin / Q000494:pharmacology
chemical_list: D000925:Anticoagulants; D003374:Coumarins; D005456:Fluorescent Dyes; D009842:Oligopeptides; D011486:Protein C; D011994:Recombinant Proteins; D018180:Thrombomodulin; C411346:benzyloxycarbonyl-glycyl--glycyl-arginine-4-methylcoumaryl-7-amide; D005340:Fibrinogen; D013925:Thromboplastin; D013917:Thrombin