Subscribe to RSS
DOI: 10.1160/TH08-10-0670
Monitoring thrombin generation: Is addition of corn trypsin inhibitor needed?
Publication History
Received:
15 October 2008
Accepted after major revision:
09 March 2009
Publication Date:
24 November 2017 (online)
Summary
Thrombin generation monitoring has the potential to be used as a clinical diagnostic tool in the near future. However, robust pre-analytical conditions may be required, and one factor that has been reported is in-vitro contact activation that might influence in-vitro measurements of thrombin generation and thereby act as an unpredictable pre-analytical variable. The aim of the current study was to investigate the influence of contact activation and the necessity of corn trypsin inhibitor (CTI) to abolish contact activation in thrombin generation measurements at low tissue factor (TF) concentrations. Thrombin generation was performed using the calibrated automated thrombinoscopy (CAT), thereby determining the endogenous thrombin potential (ETP), peak height, and the lag time, in plasma obtained from healthy volunteers. Addition of CTI after plasma preparation had no significant influence on thrombin generation triggered with 0.5 pM TF or higher, as demonstrated by unaltered ETP and lag time values between analyses with and without CTI. Addition of CTI before blood collection reduced thrombin generation triggered with 0.5 pM TF: both the ETP and peak height were significantly reduced compared to no CTI addition. In contrast, thrombin generation remained unaltered at a 1 pM TF trigger or above. This study demonstrates that addition of CTI after plasma separation is not necessary when triggering with TF concentrations of 0.5 pM and higher. Furthermore, it was demonstrated that it is not needed to pre-fill blood collecting tubes with CTI when measuring thrombin generation at TF concentrations of ≥1 pM.
-
References
- 1 Spronk HM, Govers-Riemslag JW, ten Cate H. The blood coagulation system as a molecular machine. Bioessays 2003; 25: 1220-1228.
- 2 Ten Cate-Hoek AJ, Dielis AW, Spronk HM. et al. Thrombin generation in patients after acute deep-vein thrombosis. Thromb Haemost 2008; 100: 240-245.
- 3 Hron G, Kollars M, Binder BR. et al. Identification of patients at low risk for recurrent venous thromboembolism by measuring thrombin generation. J Am Med Assoc 2006; 296: 397-402.
- 4 Regnault V, Hemker HC, Wahl D. et al. Phenotyping the haemostatic system by thrombography--potential for the estimation of thrombotic risk. Thromb Res 2004; 114: 539-545.
- 5 Dargaud Y, Beguin S, Lienhart A. et al. Evaluation of thrombin generating capacity in plasma from patients with haemophilia A and B. Thromb Haemost 2005; 93: 475-480.
- 6 Dielis AW, Balliel WM, van Oerle R. et al. Thrombomodulin-modified thrombin generation after in vivo recombinant factor VIII treatment in severe hemophilia A. Haematologica 2008; 93: 1351-1357.
- 7 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.
- 8 Besser M, Baglin C, Luddington R. et al. High rate of unprovoked recurrent venous thrombosis is associated with high thrombin-generating potential in a prospective cohort study. J Thromb Haemost 2008; 6: 1720-1725.
- 9 Eichinger S, Hron G, Kollars M. et al. Prediction of recurrent venous thromboembolism by endogenous thrombin potential and D-dimer. Clin Chem 2008; 54: 2042-2048.
- 10 Dargaud Y, Luddington R, Gray E. et al. Effect of standardization and normalization on imprecision of calibrated automated thrombography: an international multicentre study. Br J Haematol 2007; 139: 303-309.
- 11 Spronk HM, Dielis AW, De Smedt E. et al. Assessment of thrombin generation II: Validation of the Calibrated Automated Thrombogram in platelet-poor plasma in a clinical laboratory. Thromb Haemost 2008; 100: 362-364.
- 12 van Veen JJ, Gatt A, Cooper PC. et al. Corn trypsin inhibitor in fluorogenic thrombin-generation measurements is only necessary at low tissue factor concentrations and influences the relationship between factor VIII coagulant activity and thrombogram parameters. Blood Coagul Fibrinolysis 2008; 19: 183-189.
- 13 Luddington R, Baglin T. Clinical measurement of thrombin generation by calibrated automated thrombography requires contact factor inhibition. J Thromb Haemost 2004; 2: 1954-1959.
- 14 Dielis AW, Castoldi E, Spronk HM. et al. Coagulation factors and the protein C system as determinants of thrombin generation in a normal population. J Thromb Haemost 2008; 6: 125-131.
- 15 Govers-Riemslag JW, Smid M, Cooper JA. et al. The plasma kallikrein-kinin system and risk of cardiovascular disease in men. J Thromb Haemost 2007; 5: 1896-1903.
- 16 Rand MD, Lock JB, van’t Veer C. et al. Blood clotting in minimally altered whole blood. Blood 1996; 88: 3432-3445.