Haemostasis pp 321-333 | Cite as

Nijmegen-Bethesda Assay to Measure Factor VIII Inhibitors

  • Elizabeth Duncan
  • Margaret Collecutt
  • Alison Street
Part of the Methods in Molecular Biology book series (MIMB, volume 992)


Hemophilia A is an inherited bleeding disorder caused by a deficiency of factor VIII coagulant activity (FVIII:C). Patients are treated with infusions of either plasma-derived or recombinant factor VIII. However, some patients develop inhibitory antibodies (inhibitors) to infused factor VIII which render it ineffective. The original Bethesda method was developed to standardize measurement of inhibitors in a factor VIII neutralization assay. One Bethesda unit is defined as that amount of inhibitor that results in 50% residual FVIII:C activity of a defined test mixture. In the Nijmegen modification of the original Bethesda method, the pH and the protein concentration of the test mixture is further standardized. As a result, the FVIII:C in the test mixture is less prone to artifactual deterioration and the test has improved specificity. Even with a standardized procedure a number of factors can affect the performance of the test and it is important for laboratory staff to be aware of their impact on the result outcome.

Key words

Factor VIII Hemophilia Coagulation Inhibitor 


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Copyright information

© Humana Press 2013

Authors and Affiliations

  • Elizabeth Duncan
    • 1
  • Margaret Collecutt
    • 2
  • Alison Street
    • 3
  1. 1.Division of HaematologyInstitute of Medical and Veterinary ScienceAdelaideAustralia
  2. 2.Alfred Pathology ServiceThe Alfred HospitalMelbourneAustralia
  3. 3.Department of HaematologyThe Alfred HospitalMelbourneAustralia

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