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Thromb Haemost 1995; 73(02): 247-251
DOI: 10.1055/s-0038-1653759
Original Article
Coagulation
Schattauer GmbH Stuttgart

The Nijmegen Modification of the Bethesda Assay for Factor VIII:C Inhibitors: Improved Specificity and Reliability

Bert Verbruggen
1   The Central Laboratory for Haematology, University Hospital St. Radboud, Nijmegen, The Netherlands
,
Irena Novakova
1   The Central Laboratory for Haematology, University Hospital St. Radboud, Nijmegen, The Netherlands
,
Hans Wessels
1   The Central Laboratory for Haematology, University Hospital St. Radboud, Nijmegen, The Netherlands
,
Jan Boezeman
1   The Central Laboratory for Haematology, University Hospital St. Radboud, Nijmegen, The Netherlands
,
Marijke van den Berg
2   The Van Creveld Clinic, University Hospital Utrecht, Utrecht, The Netherlands
,
Eveline Mauser-Bunschoten
2   The Van Creveld Clinic, University Hospital Utrecht, Utrecht, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 04 July 1994

Accepted after resubmission 28 October 1994

Publication Date:
09 July 2018 (online)

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Summary

Antibodies against factor VIII coagulant activity can appear in haemophiliacs who are treated with factor VIII preparations but also spontaneously in non-haemophiliacs. The Bethesda assay is the most commonly used method to detect these antibodies, but it lacks specificity especially in the lower range resulting in unreliable data.

Two modifications are proposed and tested to resolve the imperfections:

1. Buffering the normal plasma used in the assay- and control mixture with 0.1 M imidazole to pH 7.4.

Replacing the imidazole buffer in the control mixture by immunodepleted factor VIII deficient plasma.

These modifications allow better discrimination between positive and negative samples and improve reliability.

 

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