Comparison of three activated protein C resistance tests in the risk assessment of venous thrombosis in non-carriers of the factor V Leiden mutation

Felipe Guerrero; Catherine Arnaud; Francoise Nguyen; Bernard Boneu; Pierre Sié

doi:10.1160/TH05-08-0593

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2006; 95(04): 728-734
DOI: 10.1160/TH05-08-0593

New Technologies, Diagnostic Tools and Drugs

Schattauer GmbH

Comparison of three activated protein C resistance tests in the risk assessment of venous thrombosis in non-carriers of the factor V Leiden mutation

Authors

Felipe Guerrero

¹Laboratoire de Recherche sur la Thrombose, EA 2049, Laboratoire d’Hématologie, Hôpital Rangueil
Catherine Arnaud

²Laboratoire d’épidémiologie, Equipe «Méthodes en Recherche Clinique», CHU Toulouse;Toulouse cedex, France
Francoise Nguyen

¹Laboratoire de Recherche sur la Thrombose, EA 2049, Laboratoire d’Hématologie, Hôpital Rangueil
Bernard Boneu

¹Laboratoire de Recherche sur la Thrombose, EA 2049, Laboratoire d’Hématologie, Hôpital Rangueil
Pierre Sié

¹Laboratoire de Recherche sur la Thrombose, EA 2049, Laboratoire d’Hématologie, Hôpital Rangueil

Abstract

Summary

Activated protein C resistance (APCR), measured using the original assay described by Dahlbäck, is a risk factor for venous thrombosis independent of the factor V Leiden (FVL) mutation. This assay is based on the activated partial thromboplastin time (APTT) after plasma exposure to activated protein C (APC).As this assay was sensitive to numerous interferences, new assays have been developed for FVL screening. The objectives of the study were to investigate the association of second generation assays for APCR with venous thrombosis in FVL non-carriers. One hundred ninety-seven subjects with a history of venous thrombosis and 211 controls were explored using 3 APCR assays, the original APTT-based assay (test A), an APTT-based assay with factorV depleted plasma pre-dilution (test B) and a direct factorX activation-based assay with the same pre-dilution (test C).We found that subjects with results in the lowest quartile of the APTT-based assays are at increased risk, compared to those in the highest quartile (test A Odds Ratio = 6.39; 95%CI 3.23–12.63; test B OR=2.72; 95%CI 1.50–4.94). There was no significant risk increase associated with test C results. After adjusting for FVIII levels, the ORs of tests A and B were similar (test A OR=3.22; 95%CI 1.47–7.08; test B OR=3.10; 95%CI 1.54–6.21). In conclusion, APTT-based assays, but not direct factor X activation-based assays, effectively detect the risk for venous thrombosis independent of FVL. Pre-dilution in factor V depleted plasma is an effective way to directly assess the risk independent of FVIII levels.

Keywords

Venous thrombosis - activated protein C resistance - factor V Leiden - factor VIII

Full Text

References

References
1 Dahlback B, Carlsson M, Svensson PJ. 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 USA 1993; 90: 1004-8.
2 Koster T, Rosendaal FR, de Ronde H. et al. Venous thrombosis due to poor anticoagulant response to activated protein C: Leiden thrombophilia study. Lancet 1993; 342: 1503-6.
3 Bertina RM, Koeleman BPC, Koster T. et al. Mutation in blood coagulation factor V associated with resistance to activated protein C. Nature 1994; 369: 64-7.
4 Cumming AM, Tait RC, Fildes S. et al. Development of resistance to activated proteinC during pregnancy. Br J Haematol 1995; 90: 725-7.
5 Olivieri O, Friso S, Manzato F. et al. Resistance to activated protein C in healthy women taking oral contraceptives. Br J Hematol 1995; 91: 465-70.
6 Henkens CMA, Bom VJJ, Van der Meer J. Lowered APC sensitivity ratio related to increased factor VIII clotting activity. Thromb Haemost 1995; 74: 1198-9.
7 Kamphuisen PW, Eikenboom JC, Bertina RM. Elevated factor VIII levels and the risk of thrombosis. Arterioscler Thromb Vasc Biol 2001; 21: 731-8.
8 Ehrenforth S, Radkte KP, Scharrer L. Acquired activated proteinC resistance in patients with lupus anticoagulants. Thromb Haemost 1995; 74: 797-8.
9 Yamazaki T, Nicolas G, Sorensen K. et al. Molecular basis of quantitative factor V deficiency associated with factor V R2 haplotype. Blood 2002; 100: 2515-21.
10 Castoldi E, Brugge J, Nicolas G. et al. Impaired APC cofactor activity of factor V plays a major role in the APC resistance associated with the factor V Leiden (R 506Q) and R2 (Hl 299R) mutations. Blood 2004; 103: 4173-9.
11 Souto JC, Almasy L, Borrell M. et al. Genetic determinants of hemostasis phenotypes in Spanish families. Circulation 2000; 101: 1546-51.
12 Soria J, Almasy L, Souto J. et al. A new locus on chromosome 18 that influences normal variation in activated protein C resistance phenotype and factor VIII activity and its relation to thrombosis susceptibility. Blood 2003; 101: 163-7.
13 Rodeghiero F, Tosetto A. Activated proteinC resistance and factor V Leiden mutation are independent risk factors for venous thromboembolism. Ann Int Med 1999; 130: 643-50.
14 De Visser MC, Rosendaal FR, Bertina RM. A reduced sensitivity for activated proteinC in the absence of factor V Leiden increases the risk of venous thrombosis. Blood 1999; 93: 1271-6.
15 Graf LL, Welsh CH, Qamar Z. et al. Activated protein C resistance assay detects thrombotic risk factors other than factor V Leiden. Am J Clin Pathol 2003; 119: 52-60.
16 Svensson PJ, Zöller B, Dahlbäck B. Evaluation of original and modified APCresistance tests in unselected outpatients with clinically suspected thrombosis and in healthy controls. Thromb Haemost 1997; 77: 332-5.
17 Shen L, Dahlback B. Factor V and proteinS as synergistic cofactors to activated proteinC in degradation of factor Vllla. J Biol Chem 1994; 269: 18735-8.
18 Polack B, Schved JF, Boneu B. Groupe d'Etude sur l'Hemostase et la Thrombose' (GEHT). Preanalytical recommendations of the 'Groupe d'Etude sur l'Hemostase et la Thrombose' (GEHT) for venous blood testing in hemostasis laboratories. Haemostasis 2001; 31: 61-8.
19 Sie P, Caron C, Azam J. et al. Reassessment of von Willebrand factor (VWF), VWF propeptide, factor VIII:C and plasminogen activator inhibitors 1 and 2 during normal pregnancy. Br J Haematol 2003; 121: 897-903.
20 Wells PS, Langlois NJ, Webster MA. et al. Elevated factor VIII is a risk factor for idiopathic venous thromboembolism in Canada – Is it necessary to definea new upper reference range for factor VIII?. Thromb Haemost 2005; 93: 842-6.
21 Folsom AR, Cushman M, Tsai MY. et al. A prospective study of venous thromboembolism in relation to factor V Leiden and related factors. Blood 2002; 99: 2720-5.
22 Rosendaal FR, van der Meer FJM, Visser Th. et al. ProC® Global screening test and the risk of thrombosis. Thromb Haemost. 1999 82. Suppl 731.
23 Grand'Maison A, Bates SM, Johnston M. et al. ProC Global“: a functional screening test that predicts recurrent venous thromboembolism. Thromb Haemost 2005; 93: 600-4.
24 de Ronde H, Bertina RM. Laboratory diagnosis of APCresistance: a critical evaluation of the test and the development of diagnostic criteria. Thromb Haemost 1994; 72: 880-6.
25 Visser MCH, van Hyclama Vlieg A, Tans G. et al. Determinants of the APTTand ETP-based APC sensitivity tests. J Thromb Haemost 2005; 03: 1488-94.
26 Tripodi A, Negri B, Bertina R. et al. Screening for the FV: Q 506 mutationevaluation of thirteen plasmabased methods for their diagnostic efficacy in comparison with DNA analysis. Thromb Haemost 1997; 77: 436-9.
27 Favaloro EJ, Orsag I, Bukuya M. et al. A 9-year retrospective assessment of laboratory testing for activated protein C resistance: evolution of a novel approach to thrombophilia investigations. Pathology 2002; 34: 348-55.
28 Favaloro EJ, Mirochnik O, McDonald D. Functional activated proteinC resistance assays: correlation with factor V DNA analysis is better with RVVT-than APTT-based assays. Br J Biomed Sci 1999; 56: 23-33.
29 Jennings I, Kitchen S, Woods TA. et al. Normalization does not improve between-laboratory agreement but may improve specificity of some assays for activated protein C resistance. Blood Coagul Fibrinolysis 1999; 10: 451-3.
30 Tripodi A, Chantarangkul V, Negri B. et al. Standardization of the APC resistance test. Effects of normalization of results by means of pooled normal plasma. Thromb Haemost 1998; 79: 564-6.