The Effect of Va rious Anticoagulant/Anti platelet Mixtures on Determination of Plasminogen Activator Inhibitor, Platelet Proteins and Hemostasis Farameters

Summary

Blood collected in different anticoagulant/antiplatelet agents(ETP, EDTA, citrate, citratelcitric acid pH 4.5 and CTAD) was compared with respect to determination of PAI-1 activity and PAI-1 antigen. βTG and PF4 were analysed as markers of platelet release. Both the middle layer and the remaining layer of the plasma were studied. Moreover vWF:Ag, FVII:Ag, ECLI t-PA:Ag, t-PA activity, APTT VIII: C and VII: C were assayed in blood collected in citrate and CTAD

PAI-1 activity showed the same level in all citrate based anticoagulants and ETP and no increase was found in blood standing for 2 hours at room temperature. On the contrary quick handling was most important for determination of PAI-1 antigen. In tubes anticoagulated with citrate no significant increase was found if the sample was prepared within 1 hour. EDTA was not suitable as anticoagulant mixture. Tubes containing the antiplatelet mixture CTAD could be used for determination of PAI activity, PAI antigen, vWF:Ag, FVIIiAg, t-PA activity and APTT.

For measurement of PAI-1 antigen quick handling of blood anticoagulated with antiplatelet mixtures are preferable, and plasma treated in that manner could also be used to assay some hemostasis parameters.

Keywords

Blood sampling - Plasminogen activator inhibitor - Platelets - β-Thromboglobulin - Platelet factor 4

• Reference

• 1 Sprengers ED, Kluft C. Plasminogen activator inhibitors. Blood 1987; 69: 381-387
• 2 Kruithof EK O. Plasminogen activator inhibitor type 1 Biochemical, biological and clinical aspects. Fibrinolysis 1988; 2: 59-70
• 3 De Clerck PJ, Alessi MC, Verstreken M, Kruithof EK O, Juhan-Vague I. Measurement of plasminogen activator inhibitor (PAI-1) in plasma with an ELISA based on two murine monoclonal antibodies. Thromb Haemostas 1987; 58: 447 Abstr 1650
• 4 Juhan-Vague I, Alessi MC, Fossat C, Declerck PJ, Kruithof EK O. Plasma determination of plasminogen activator inhibitor 1 antigen must be performed in blood collected on antiplatelet/anticoagulant mixture. Thromb Haemostas 1987; 58: 1096
• 5 Ludlam CA, Cash JD. Studies of the liberation of p-thromboglobulin from human platelet in vitro. Br J Haematol 1976; 33: 239-247
• 6 Amiral J, Trebaol JF, Adam M. Elisa evaluation of beta thrombo-globulin and platelet factor 4. Proc IVth Int Meeting of Danubian League against Thrombosis and Hemorrhagic Diseases; Istanbul: 1985. pp 376-382
• 7 Råsi V. ²-thromboglobulin in plasma: False high values caused by platelet enrichment of the top layer of plasma during centrifugation. Thromb Res 1979; 15: 543-552
• 8 Eriksson E, Rånby M, Gyzander E, Risberg B. Determination of plasminogen activator inhibitor in plasma using t-PA and a chromogenic single point poly-D-lysine stimulated assay. Thromb Res 1988; 50: 91-101
• 9 Kruithof EK O, Nicoloso G, Bachmann F. Plasminogen activator inhibitor 1. Development of a radioimmunoassay and observations on its plasma concentration during venous occlusion and after platelet aggregation Blood 1987; 70: 1645-1653
• 10 Eriksson E, Risberg B. Measurement of tissue plasminogen activator in plasma. A comparison of 3 methods and description of a new improved technique Thromb Res 1987; 46: 213-223
• 11 Nilsson IM. aemorrhagic and Thrombotic Diseases. John Wiley and Sons; London: 1974
• 12 Ingerslev I. A sensitive ELISA for von Willebrand factor (vWf:Ag). Scand J Clin Lab Invest 1987; 47: 143-149
• 13 Woolf CM. Principles of Biometry. D. van Norstrand Inc; New York: 1968
• 14 Gyzander E, Eriksson E, Teger-Nilsson A-C. A sensitive assay for t-PA activity in plasma using adsorption on lysine sepharose. Thromb Res 1984; 35: 547-558
• 15 Kaplan KL, Owen J. Plasma levels of p-thromboglobulin and platelet factor 4 as indices of platelet activation in vivo. Blood 1981; 57: 199-202
• 16 Booth NA, Simpson AJ, Croll A, Bennet B, Mac Gregor IR. Plasminogen activator inhibitor (PAI-1) in plasma and platelets. Br J Haematol 1988; 70: 327-333