Thrombin generation assay using factor IXa as a trigger to quantify accurately factor VIII levels in haemophilia A

J Thromb Haemost. 2011 Aug;9(8):1549-55. doi: 10.1111/j.1538-7836.2011.04358.x.

Abstract

Background: The available methods for measuring factor VIII (FVIII) activity suffer reportedly from lack of sensitivity and precision in the < 1 IU dL(-1) range. This precludes correlation of clinical phenotype with FVIII levels.

Objectives: To study a possible association between clinical phenotype in patients with FVIII levels < 1 IU dL(-1).

Methods/results: The FIXa-driven FVIII assay (FVIII-CAT) has a detection limit of 0.05 IU dL(-1). For the range of 0-2 IU dL(-1) FVIII, the intra-assay coefficient of variation (CV) is around 2% and the inter-assay CV is about 8%. We tested 30 hemophiliacs with FVIII:C between < 1 and 6 IU dL(-1) as measured in the one-stage clotting assay using the FVIII-CAT assay. For genetic defects related to moderate hemophilia, the FVIII-CAT test finds FVIII levels that are in good agreement with those determined with the one-stage assay. Of the 21 hemophilic patients with FVIII < 1 IU dL(-1), four patients exhibited a mild bleeding phenotype. When we applied TF-initiated thrombin generation, patients with a mild clinical phenotype showed significantly higher endogenous thrombin potentials.

Conclusion: The novel developed FVIII assay measures accurately FVIII levels below 1 IU dL(-1). Its application demonstrated that the clinical heterogeneity in individuals with < 1 IU dL(-1) FVIII is not associated with their FVIII level.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Biomarkers / blood
  • Blood Coagulation Tests*
  • Factor VIII / analysis*
  • Factor Xa / metabolism*
  • France
  • Hemophilia A / blood
  • Hemophilia A / diagnosis*
  • Humans
  • Kinetics
  • Middle Aged
  • Netherlands
  • Phenotype
  • Predictive Value of Tests
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Severity of Illness Index
  • Thrombin / metabolism*
  • Thromboplastin / metabolism
  • Young Adult

Substances

  • Biomarkers
  • Factor VIII
  • Thromboplastin
  • Thrombin
  • Factor Xa