Thrombin generation in plasma of healthy adults and children: Chromogenic versus fluorogenic thrombogram analysis

Katrien Devreese; Walter Wijns; Isabelle Combes; Soetkin Vankerckhoven; Marc F. Hoylaerts

doi:10.1160/TH07-03-0210

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2007; 98(03): 600-613
DOI: 10.1160/TH07-03-0210

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Thrombin generation in plasma of healthy adults and children: Chromogenic versus fluorogenic thrombogram analysis

Katrien Devreese

¹Coagulation Laboratory, Department of Clinical Chemistry, Microbiology and Immunology, Ghent University Hospital, Ghent, Belgium

,

Walter Wijns

²Haemostasis Laboratory, Department of Clinical Pathology, Hôpital Erasme, Free University of Brussels, Brussels, Belgium

,

Isabelle Combes

¹Coagulation Laboratory, Department of Clinical Chemistry, Microbiology and Immunology, Ghent University Hospital, Ghent, Belgium

,

Soetkin Vankerckhoven

³Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium

,

Marc F. Hoylaerts

³Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium

› Author Affiliations

Abstract

Summary

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 standardizedTF 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.

Keywords

Tissue factor - coagulation assays - endogenous thrombin potential - fibrinogen

Full Text

References

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