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Thromb Haemost 2015; 114(04): 804-811
DOI: 10.1160/TH14-12-1062
New Technologies, Diagnostic Tools and Drugs
Schattauer GmbH

Diagnostic accuracy study of a factor VIII ELISA for detection of factor VIII antibodies in congenital and acquired haemophilia A

Paul Batty
1   The Royal London Hospital Haemophilia Centre, Barts and The London School of Medicine & Dentistry, QMUL, London, UK
,
Gary W. Moore
2   Haemostasis and Thrombosis, Viapath Guy’s and St Thomas’ NHS Foundation Trust, London, UK
,
Sean Platton
3   The Royal London Hospital Haemophilia Centre, Barts Health NHS Trust, London, UK
,
James C. Maloney
2   Haemostasis and Thrombosis, Viapath Guy’s and St Thomas’ NHS Foundation Trust, London, UK
,
Ben Palmer
4   The United Kingdom National Haemophilia Database, Manchester, United Kingdom
,
Louise Bowles
3   The Royal London Hospital Haemophilia Centre, Barts Health NHS Trust, London, UK
,
John K. Pasi
1   The Royal London Hospital Haemophilia Centre, Barts and The London School of Medicine & Dentistry, QMUL, London, UK
,
Savita Rangarajan
5   Centre for Haemostasis and Thrombosis, Guy’s and St Thomas’ NHS Foundation Trust, London, UK
,
Daniel P. Hart
1   The Royal London Hospital Haemophilia Centre, Barts and The London School of Medicine & Dentistry, QMUL, London, UK
› Author Affiliations Financial support: Paul Batty has received an unrestricted research grant from Octapharma.
Further Information

Publication History

Received: 19 December 2014

Accepted after major revision: 19 May 2015

Publication Date:
29 November 2017 (online)

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Summary

Antibody formation to factor VIII (FVIII) remains the greatest clinical and diagnostic challenge to the haemophilia-treating physician. Current guidance for testing for inhibitory FVIII antibodies (inhibitors) recommends the functional Nijmegen-Bethesda assay (NBA). A FVIII ELISA offers a complementary, immunological approach for FVIII antibody testing. It was the aim of this study to retrospectively evaluate the performance of a FVIII ELISA (index) for detection of FVIII antibodies, compared with the NBA (reference). All samples sent for routine FVIII antibody testing at two haemophilia Comprehensive Care Centres, were tested in parallel using the NBA and a solid-phase, indirect FVIII ELISA kit (Immucor). A total of 497 samples from 239 patients (severe haemophilia A=140, non-severe haemophilia A=85, acquired haemophilia A=14) were available for analysis. Sixty-THree samples tested positive by the NBA (prevalence 12.7 %, 95 % confidence interval [CI], 9.9–15.9 %), with a median inhibitor titre of 1.2 BU/ml (range 0.7–978.0). The FVIII ELISA demonstrated a specificity of 94.0 % (95 %CI, 91.3–96.0), sensitivity of 77.8 % (95 %CI, 65.5–87.3), negative predictive value of 96.7 % (95 %CI, 94.5–98.2), positive predictive value 65.3 % (95 %CI, 53.5–76.0), negative likelihood ratio 0.2 (95 %CI, 0.1–0.4), positive likelihood ratio 13.0 (95 %CI, 8.7–19.3) and a diagnostic odds ratio of 54.9 (95 %CI, 27.0–112.0). Strong positive correlation (r=0.77, p< 0.001) was seen between the results of the NBA (log adjusted) and FVIII ELISA optical density. In conclusion, FVIII ELISA offers a simple, specific, surveillance method enabling batch testing of non-urgent samples for the presence of FVIII antibodies.

Keywords

Factor VIII inhibitors - haemophilia A - ELISA - Bethesda - sensitivity

 

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