Clot waveform analysis in patients with haemophilia A

Summary

Clot waveform analysis extends the interpretation of aPTT measurement curves. The curve is mathematically processed to obtain information about fibrin formation kinetics including semiquantitative determination of thrombin, prothrombinase and tenase activity. Patients, method: In this study the feasibility of clot waveform analysis for monitoring of haemophilia A was investigated using blood samples from healthy controls as well as haemophilia A patients under various clinical conditions. Results: Thrombin, prothrombinase and tenase activity show a high correlation to factor VIII levels. Tenase activity was found to exhibit a linear relationship to factor VIII levels over a very large concentration range and was able to discriminate patients with severe, moderate and mild haemophilia. Conclusion: Clot waveform analysis is an easy, fast and cheap method to access disturbances in clot formation and can be done without any additional measurements beside an aPTT.

Zusammenfassung

Clot-Waveform-Analyse leitet zusätzlich Daten aus aPTT-Messungen aus. Die aufgezeichnete Messkurve wird mathematisch aufbereitet und die Kinetik der Fibrinbildung einschließlich Thrombin-, Prothrombinaseund Tenaseaktivität semiquantitativ bestimmt. Patienten, Methode: Proben gesunder Blutspender als auch von Hämophilie-A-Patienten in verschiedenen klinischen Situationen wurden untersucht, um die Anwendbarkeit der Clot-Waveform-Analyse für die Überwachung der Hämophilie A zu prüfen. Ergebnisse: Thrombin-, Prothrombinaseund Tenaseaktivität zeigen eine hohe Korrelation zum Faktor-VIII-Spiegel. Ein linearer Zusammenhang wurde zwischen Tenaseaktivität und Faktor VIII-Spiegel über einen großen Konzentrationsbereich gefunden. Patienten mit schwerer, moderater oder milder Hämophilie können gut diskriminiert werden. Schlusssfolgerung: Clot-Waveform-Analyse ist eine einfache, schnelle und preiswerte Methode, um Störungen der Fibrinbildung zu untersuchen. Die Daten können aus einer aPTT-Bestimmung gewonnen werden.

Keywords

Activated partial thromboplastin time - fibrin formation kinetics - thrombin - prothrombinase - tenase - haemophilia

Schlüsselwörter

Aktivierte partielle Thromboplastinzeit - Fibrinbildungskinetik - Thrombin - Prothrombinase - Tenase - Hämophilie

• References

• 1 Braun PJ, Givens TB, Stead AG. et al. Properties of optical data from activated partial thromboplastin time and prothrombin time assays. Thromb Haemost 1997; 78: 1079-1087.
• 2 Shima M, Matsumoto T, Fukuda K. et al. The utility of activated partial thromboplastin time (aPTT) clot waveform analysis in the investigation of hemophilia A patients with very low levels of factor VIII activity (FVIII:C). Thromb Haemost 2002; 87: 436-441.
• 3 Kasuda S, Tanaka I, Shima M. et al. Effectiveness of factor VIII infusions in haemophilia A patients with high responding inhibitors. Haemophilia 2004; 10: 341-346.
• 4 Toh CH, Samis J, Downey C. et al. Biphasic transmittance waveform in the APTT coagulation assay is due to the formation of a Ca⁺⁺-dependent complex of C-reactive protein with very-low-density lipoprotein and is a novel marker of impending disseminated intravascular coagulation. Blood 2002; 100: 2522-2529.
• 5 Hoffman M, Monroe 3rd DM. A cell-based model of hemostasis. Thromb Haemost 2001; 85: 958-965.
• 6 Siegemund T, Petros S, Siegemund A. et al. Thrombin generation in severe haemophilia A and B: the endogenous thrombin potential in platelet-rich plasma. Thromb Haemost 2003; 90: 781-786.
• 7 Wolberg AS, Gabriel DA, Hoffman M. Analyzing fibrin clot structure using a microplate reader. Blood Coagul Fibrinol 2002; 13: 533-953.
• 8 Chatterjee MS, Denney WS, Jing H, Diamond SL. Systems biology of coagulation initiation: kinetics of thrombin generation in resting and activated human blood. PLoS Comput Biol. 2010 06..
• 9 Hockin MF, Jones KC, Everse SJ, Mann KG. A model for the stoichiometric regulation of blood coagulation. J Biol Chem 2002; 277: 18322-18333.
• 10 Savitzky A, Golay MJE. Smoothing and differentiation of data by simplified least squares procedures. Anal Chem 1964; 36: 1627.
• 11 Rand MD, Lock JB, van’t Veer C. et al. Blood clotting in minimally altered whole blood. Blood 1996; 88: 3432-3445.
• 12 Higgins DL, Lewis SD, Shafer JA. Steady state kinetic parameters for the thrombin-catalyzed conversion of human fibrinogen to fibrin. J Biol Chem 1983; 258: 9276-9282.