Thromb Haemost 1994; 72(02): 255-60
DOI: 10.1055/s-0038-1648849
Original Article
Schattauer GmbH Stuttgart

Multicenter Evaluation of a Kit for Activated Protein C Resistance on Various Coagulation Instruments Using Plasmas from Healthy Individuals

S Rosén
Chromogenix AB, Mölndal, Sweden and Department of Clinical Chemistry and Department of Coagulation Disorders, University of Lund, Malmö General Hospital, Malmö, Sweden
,
K Johansson
Chromogenix AB, Mölndal, Sweden and Department of Clinical Chemistry and Department of Coagulation Disorders, University of Lund, Malmö General Hospital, Malmö, Sweden
,
K Lindberg
Chromogenix AB, Mölndal, Sweden and Department of Clinical Chemistry and Department of Coagulation Disorders, University of Lund, Malmö General Hospital, Malmö, Sweden
,
B Dahlbäck
Chromogenix AB, Mölndal, Sweden and Department of Clinical Chemistry and Department of Coagulation Disorders, University of Lund, Malmö General Hospital, Malmö, Sweden
,
(for the APC resistance study group) › Author Affiliations
Further Information

Publication History

Received: 14 January 1994

Accepted after revision05 April 1994

Publication Date:
24 July 2018 (online)

Summary

Recently a new hemostatic disorder has been described which appears to be an important risk factor for familial thromboembolism. The disorder is characterized by a poor anticoagulant response to activated Protein C (APC) and has been shown to be due to lack of an APC cofactor activity which is a property of factor V.

A kit for determining the response of plasma samples towards addition of APC in an APTT-based assay - COATEST APC Resistance -has been evaluated on 35 coagulation instruments in a multicenter study involving 32 laboratories. A lyophilized normal plasma and identical plasma aliquots from 20 individuals, one of whom had a borderline resistance to APC, were analysed in each laboratory and the sensitivity of each plasma to APC was determined as the ratio between the clotting times obtained in the presence and absence of APC (APC ratio).

The plasma from the individual with a borderline resistance to APC activity was correctly classified as the lowest responder in each laboratory, with an APC ratio in the range 1.6-2.4. In comparison, plasmas from individuals with a pronounced response to APC activity resulted in APC ratios above 3.4 in most cases. Interestingly, although the actual APT time for a plasma from a given individual showed a more than 10 s difference due to the type of instrumentation used, the variation in the APC ratio was limited. A similar discrimination was also obtained from evaluation of the actual prolongation of the clotting time in the presence of APC.

The intra-laboratory coefficient of variation for the clotting times were on average 2.0% and 3.9% in the absence and presence of APC, respectively, indicating that the precision for the prolonged clotting times obtained in the presence of APC is sufficient to allow a safe assignment of the APC response. The APC ratio for the lyophilized normal plasma was 2.7 ± 0.2 (2 S.D.) illustrating a narrow distribution between instruments which shows the feasibility of including such plasma for assay validation. Altogether, the results indicate that all the coagulation instruments included in the study can be used for detection of individuals with resistance to APC activity through determination of the APC ratio or the prolongation time.

 
  • References

  • 1 Egeberg O. Inherited antithrombin III deficiency causing thrombophilia. Thromb Diathesis Haemorrhagica 1965; 13: 516-530
  • 2 Thaler E, Lechner K. Antithrombin III deficiency and thromboembolism. Clin Haematol 1981; 10: 369-390
  • 3 Griffin J, Evatt B, Zimmermann T, Kleiss A, Wideman C. Deficiency of Protein C in congenital thrombotic disease. J Clin Invest 1981; 68: 1370-1373
  • 4 Broekmans AW, Veltkamp J, Bertina R. Congenital Protein C deficiency and venous thromboembolism: a study in three Dutch families. N Engl J Med 1983; 309: 340-344
  • 5 Comp P, Nixon R, Cooper M, Esmon C. Familial Protein S deficiency is associated with recurrent thrombosis. J Clin Invest 1984; 74: 2082-2088
  • 6 Schwartz H, Fischer P, Batard M, Griffin J. Plasma Protein S deficiency in familial thrombotic disease. Blood 1984; 64: 1297-1300
  • 7 Mannucci PM, Tripodi A. Inherited factors in thrombosis. Blood Rev 1988; 2: 27-35
  • 8 Gladson CL, Scharrer I, Hach V, Beck KH, Griffin J. The frequency of type I heterozygous Protein S and Protein C deficiency in 141 unrelated young patients with venous thrombosis. Thromb Haemost 1988; 59: 18-22
  • 9 Heijboer H, Brandjes D, Biiller H, Sturk A, ten Cate JW. Deficiencies of coagulation-inhibiting and fibrinolytic proteins in outpatients with deep-vein thrombosis. New Engl J Med 1990; 323: 1512-1516
  • 10 Tabemo MD, Tomas JF, Alberca I, Orfao A, Borrasca AL, Vicente V. Incidence and clinical characteristics of hereditary disorders associated with venous thrombosis. Am J Hematol 1991; 36: 249-254
  • 11 Malm J, Laurell M, Nilsson IM, Dahlback B. Thromboembolic disease -critical evaluation of laboratory investigation. Thromb Haemost 1992; 68: 7-13
  • 12 Pabinger I, Bruckner S, Kyrle PA, Schneider B, Korninger HC, Niessner H, Lechner K. Hereditary deficiency of antithrombin III, Protein C and Protein S: prevalence in patients with a history of venous thrombosis and criteria for rational patient screening. Blood Coag and Fibrinol 1992; 3: 547-553
  • 13 Henkens CMA, van der Meer J, Hillege JL, van der Schaaf W, Bom VJJ, Halie MR. The clinical expression of hereditary Protein C and protein S deficiency: a relation to clinical thrombotic risk factors and to levels of Protein C and Protein S?. Blood Coag and Fibrinol 1993; 4: 555-562
  • 14 Lechner K, Pabinger-Fasching I. Lupus anticoagulants and thrombosis, a study of 25 cases and a review of the literature. Haemostasis 1985; 15: 254-262
  • 15 Triplett D. Antiphospholipid antibodies and thrombosis. Arch Pathol Lab Med 1993; 117: 78-88
  • 16 Levi M, Hack E, de Boer JP, Brandjes D, Biiller H, ten Cate JW. Reduction of contact activation related fibrinolytic activity in FXII deficient patients. J Clin Invest 1991; 88: 1155-1160
  • 17 Munkvad S, Jespersen J, Gram J, Kluft C. Depression of factor XH-depen-dent fibrinolytic activity characterizes patients with early myocardial reinfarction after tissue-type plasminogen activator therapy. J Am Coll Cardiol 1991; 18: 454-458
  • 18 Halbmayer WM, Mannhalter C, Feichtinger C, Rubi K, Fischer M. The prevalence of FXII deficiency in 103 orally anticoagulated outpatients suffering from recurrent venous and/or arterial thromboembolism. Thromb Haemost 1992; 68: 285-290
  • 19 Lammle B, Wuillemin WA, Huber I, Krauskopf M, Ziircher C, Pflugshaupt R, Furlan M. Thromboembolism and bleeding tendency in congenital factor XII deficiency - a study on 74 subjects from 14 Swiss families. Thromb Haemost 1991; 65: 117-121
  • 20 Nilsson IM, Ljungner H, Tengbom L. Two different mechanisms in patients with venous thrombosis and defective fibrinolysis. Low concentration of plasminogen activator or increased concentration of plasminogen activator inhibitor. Br Med J 1985; 290: 1453-1456
  • 21 Juhan-Vague I, Valadier J, Alessi MC, Aillaud MF, Ansaldi J, Philip-Joet C, Holvoet P, Semmadimigni A, Collen D. Deficient t-PA release and elevated PA inhibitor levels in patients with spontaneous or recurrent deep venous thrombosis. Thromb Haemost 1987; 57: 67-72
  • 22 Wiman B, Ljungberg B, Chmielewska J, Urden G, Blomback M, Johnsson H. The role of the fibrinolytic system in deep venous thrombosis. J Lab Clin Med 1985; 105: 265-270
  • 23 Jprgensen M, Bonnevie-Nielsen V. Increased concentration of the fast-acting plasminogen activator inhibitor in plasma associated with familial venous thrombosis. Br J Haematol 1987; 65: 175-180
  • 24 Nicoloso G, Hauert J, Kruithof EKO, van Melle G, Bachmann F. Discrimination d’une population normale et d’un collectif de patients ayant presente un antecedent thromboembolique idiopathique au moyen des tests fibrino-lytiques. Schweiz Med Wochenschr 1988; 118: 122-121
  • 25 Hamsten A, Wiman B, de Faire U, Blomback M. Increased plasma levels of a rapid inhibitor of tissue plasminogen activator in young survivors of myocardial infarction. N Engl J Med 1985; 33: 1557-1563
  • 26 Hach-Wunderle V, Scharrer I. Decreased t-PA activity in patients with venous thromboembolism. Thromb Haemost 1993; 69: 1053
  • 27 Ridker P, Vaughan D, Stampfer M, Manson JA, Shen C, Newcomer L, Goldhaber S, Hennekens C. Baseline fibrinolytic state and the risk of future venous thrombosis. A prospective study of endogenous tissue type plasminogen activator and plasminogen activator inhibitor. Circulation 1992; 85: 1822-1827
  • 28 Dawson S, Henney A. The status of PAI-1 as a risk factor for arterial and thrombotic disease: A review. Atherosclerosis 1992; 95: 105-117
  • 29 Dahlback B, Carlsson M, Svensson P. Familial thrombophilia due to a previously unrecognized mechanism characterized by poor anticoagulant response to activated Protein C: Prediction of a cofactor to activated Protein C. Proc Natl Acad Sci 1993; 90: 1004-1008
  • 30 Dahlback B. Familial thrombophilia associated with resistance to activated Protein C is due to deficiency of a novel Protein C cofactor. Thromb Haemost 1993; 69: 978
  • 31 Dahlback B, Hildebrand B. Inherited resistance to activated protein C is corrected by anticoagulant cofactor activity found to be a property of factor V. Proc Natl Acad Sci 1994; 91: 1396-1400
  • 32 Svensson P, Dahlback B. Novel mechanism for thrombosis characterized by poor anticoagulant response to activated Protein C constitutes a major cause of thrombophilia. Thromb Haemost 1993; 69: 999
  • 33 Svensson PJ, Dahlback B. Resistance to activated Protein C as a basis for venous thrombosis. New Engl J Med 1994; 330: 517-522
  • 34 Svensson P, Dahlback B. Twenty novel families with thrombophilia and inherited resistance to activated Protein C. Thromb Haemost 1993; 69: 1252
  • 35 Griffin J, Evatt B, Widemann C, Fernandez J. Anticoagulant Protein C pathway defective in majority of thrombophilic patients. Blood 1993; 82: 1989-1993
  • 36 Koster T, Rosendaal FR, de Ronde H, Briet E, Vandenbroucke JP, Bertina RM. Venous thrombosis due to poor anticoagulant response to activated Protein C: Leiden thrombophilia study. Lancet 1993; 342: 1503-1506
  • 37 Faioni EM, Franchi F, Asti D, Sacchi E, Bemardi F, Mannucci PM. Resistance to activated Protein C in nine thrombophilic families: Interference in a Protein S functional assay. Thromb Haemost 1993; 70: 1067-1071
  • 38 Johansson K, Lindberg K, Menschik M, Rosen S. Evaluation of a kit for detection of resistance in plasma towards activated Protein C activity in a modified APTT assay. Thromb Haemost 1993; 69: 1120