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Thromb Haemost 1985; 54(04): 818-823
DOI: 10.1055/s-0038-1660140
Original Article
Schattauer GmbH Stuttgart

Clinical Application of a Chromogenic Substrate Method for Determination of Factor VIII Activity

S Rosén
1   The KabiVitrum Hematology, The Peptide Research, Mölndal, Sweden
,
M Andersson
1   The KabiVitrum Hematology, The Peptide Research, Mölndal, Sweden
,
M Blombäck
2   The Department of Clinical Chemistry and Blood Coagulation, Karolinska Instituted Stockholm, Sweden
,
U Hégglund
2   The Department of Clinical Chemistry and Blood Coagulation, Karolinska Instituted Stockholm, Sweden
,
M J Larrieu
3   The INSERM U.143, Institut de Pathologie Cellulaire, Hôpital de Bicêtre, le Kremlin-Bicetre, France
,
M Wolf
3   The INSERM U.143, Institut de Pathologie Cellulaire, Hôpital de Bicêtre, le Kremlin-Bicetre, France
,
C Boyer
3   The INSERM U.143, Institut de Pathologie Cellulaire, Hôpital de Bicêtre, le Kremlin-Bicetre, France
,
C Rothschild
3   The INSERM U.143, Institut de Pathologie Cellulaire, Hôpital de Bicêtre, le Kremlin-Bicetre, France
,
I M Nilsson
4   The Department for Coagulation Disorders, University of Lund, Allmänna Sjukhuset, Malmö, Sweden
,
E Sjörin
4   The Department for Coagulation Disorders, University of Lund, Allmänna Sjukhuset, Malmö, Sweden
,
H Vinazze
5   The Laboratorium für Blutgerinnung, Linz, Austria
› Author Affiliations
Further Information

Publication History

Received 20 May 1985

Accepted 17 September 1985

Publication Date:
19 July 2018 (online)

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Summary

A chromogenic substrate kit for the determination of factor VIII activity (COATEST® Factor VIII) has been evaluated in five different laboratories, one of them using a semi-automated procedure. This chromogenic method was compared to one-stage clotting assays for factor VIII determination in plasmas from healthy subjects, carriers of hemophilia A, severe, mild and moderate hemophilia A as well as von Willebrand’s patients. In all these cases, a high correlation between these two methods was obtained (r = 0.96-0.99, n = 385) with a good agreement of the assigned potencies at all levels of factor VIII. A good correlation (r = 0.94) was also obtained for the levels of factor VIII after infusion of concentrates in six severe hemophiliacs or after administration of DDAVP to von Willebrand’s patients.

The chromogenic method is insensitive to preactivation of factor VIII by thrombin, thus yielding valid potency assignments also in these situations.

The precision was higher with the chromogenic method than with the one-stage clotting assays (C.V. = 2-5% vs 4-15%). Altogether, the new chromogenic substrate method has proven itself suitable for determination of factor VIII in plasma and concentrates.

Key words

Factor VIII activity assay - Chromogenic substrate kit - Clinical evaluation
 

  • References

  • 1 Seghatchian MJ, Miller-Anderson M. A colorimetric evaluation of. factor VIII :C potency. Med Lab Sci 1978; 35: 347-354
    PubMedGoogle Scholar
  • 2 Friberger P, Vinazzer H. Assay of factor VIII: C with a chromogenic substrate. In: Transactions of the first Danubesymposium on Thrombosis and Haemostasis Vinazzer H. (ed) Medicus Verlag Berlin: 1979: 117-121
    Google Scholar
  • 3 Vinazzer H. Photometric assay of factor VIII :C with a chromogenic substrate. Thromb Haemostas 1979; 42: 238 (Abstr)
    PubMedGoogle Scholar
  • 4 Rosén S, Friberger P, Andersson M, Vinazzer H. A new chromogenic assay for determination of human factor VIII: C activity. 18th Congr Int Soc Haematol and 16th Congr Int Soc Blood Transf, Montreal, Aug 16-22 1980: 1315
    PubMedGoogle Scholar
  • 5 Claeson G, Aurell L, Karlsson G, Friberger P. Substrate structure and activity relationships. Proc 16th Int Congr Haematol, Kyoto, Sept 5-11 1976
    PubMedGoogle Scholar
  • 6 Rosen S. Assay of factor VIII: C with a chromogenic substrate. Scand J Haematol 1984; 33 (Suppl. 140) 139-145
    PubMedGoogle Scholar
  • 7 Mikaelsson M, Oswaldsson U. Standardization of VIII :C assays: A manufacturer’s view. Scand J Haematol 1984; 33 (Suppl. 41) 79-86
    PubMedGoogle Scholar
  • 8 Wood WI, Capon DJ, Simonsen CC, Eaton DL, Gitschier J, Keyt B, Seebury PH, Smith DH, Hollingshead P, Wion KL, Delwart E, Tuddenham E GD, Vehar GA, Lawn RM. Expression of active human factor VIII from recombinant DNA clones. Nature 1984; 312: 330-337
    CrossrefPubMedGoogle Scholar
  • 9 Toole JJ, Knopf JL, Wozney JM, Sultzman LA, Buecker JL, Pittman DD, Kaufman RJ, Brown E, Shoemaker C, Orr EC, Amphlett GN, Foster WB, Coe ML, Knutson GJ, Fass DN, Hewich RM. Molecular cloning of a cDNA encoding human antihaemophilic factor. Nature 1984; 312: 342-347
    CrossrefPubMedGoogle Scholar
  • 10 Rosen S, Oswaldsson U, Blomback M, Larrieu M, Nilsson IM, Vinazzer H. Evaluation of a chromogenic method for determining factor VIII :C in haemophiliacs, von Willebrand patients and factor VIII concentrates. XVth World Fed Hemophilia Congr, Stockholm June 27-July 1 1983 Abstract no 241
    PubMedGoogle Scholar
  • 11 Rosén S, Oswaldsson U, Blombäck M, Larrieu M, Nilsson IM, Vinazzer H. A collaborative comparison of a chromogenic factor VIII: C method with one- and two-stage methods. Thromb Haemostas 1983; 50: 112 (Abstr)
    PubMedGoogle Scholar
  • 12 Nilsson IM, Blombäck M, von Francken I. On an inherited autosomal diathesis with antihaemophilic globulin (AHG) deficiency and prolonged bleeding time. Acta Med Scand 1957; 159: 35-37
    PubMedGoogle Scholar
  • 13 Hardisty RM, Macpherson JC, Ingram GIC. A one-stage factor VIII (antihaemophilic globulin) assay and its use on venous and capillary plasma. Thrombos Diathes Haemorrh 1962; 7: 215-229
    PubMedGoogle Scholar
  • 14 Langdell RC, Wagner RH, Brinkhous KM. Effect of antihemophilic factor on one-stage clotting tests. J Lab Clin Med 1953; 41: 637-647
    PubMedGoogle Scholar
  • 15 Nilsson IM. Haemorrhagic and thrombotic diseases. John Wiley & Sons, London 1974
    PubMedGoogle Scholar
  • 16 Nilsson IM, Kirkwood TB L, Barrowcliffe TW. In vivo recovery of factor VIII: A comparison of one-stage and two-stage assay methods. Thromb Haemostas 1979; 42: 1230-1239
    PubMedGoogle Scholar
  • 17 Proctor RR, Rapaport SI. The partial thromboplastin time with kaolin. A simple screening test for first stage plasma clotting deficiencies. Amer J Clin Pathol 1961; 36: 212-219
    PubMedGoogle Scholar
  • 18 Barrowcliffe TW, Tydeman MS, Kirkwood TB L, Thomas DP. Standardization of factor VIII-III. Establishment of a stable reference plasma for factor VUI-related activities. Thromb Haemostas 1983; 50: 690-696
    PubMedGoogle Scholar
  • 19 Nilsson IM. In memory of Erik Jorpes - von Willebrand’s Disease from 1926-1983. Scand J Haematol 1984; 33 (Suppl. 40) 21-43
    PubMedGoogle Scholar
  • 20 Armitage P. Statistical Methods in Medical Research. 4th ed (1981). Blackwell Scientific Publications; Oxford, Great Britain: 1981
  • 21 Österud B, Rapaport SI, Schiffman S, Chong MY. Formation of intrinsic factor X-activator activity with special reference to the role of thrombin. Br J Haematol 1971; 21: 643-660
    CrossrefPubMedGoogle Scholar
  • 22 Hoyer LW, Trabold NC. The effect of thrombin on human factor VIII. Cleavage of the factor VIII procoagulant protein during activation. J Lab Clin Med 1981; 97: 50-64
    PubMedGoogle Scholar