Comparison of Clot-based, Chromogenic, and Fluorescence Assays for Measurement of Factor VIII Inhibitors in the U.S. Hemophilia Inhibitor Research Study

Connie H Miller; Anne S Rice; Brian Boylan; Amy D Shapiro; Steven R Lentz; Brian M Wicklund; Fiona M Kelly; J. Michael Soucie; Hemophilia Inhibitor Research Study Investigators

doi:10.1111/jth.12259

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Comparison of Clot-based, Chromogenic, and Fluorescence Assays for Measurement of Factor VIII Inhibitors in the U.S. Hemophilia Inhibitor Research Study

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Comparison of Clot-based, Chromogenic, and Fluorescence Assays for Measurement of Factor VIII Inhibitors in the U.S. Hemophilia Inhibitor Research Study

Connie H Miller, Anne S Rice, Brian Boylan, Amy D Shapiro, Steven R Lentz, Brian M Wicklund, Fiona M Kelly, J. Michael Soucie and Hemophilia Inhibitor Research Study Investigators

Journal of thrombosis and haemostasis, Vol.11(7), pp.1300-1309

07/2013

DOI: 10.1111/jth.12259

PMCID: PMC4477744

PMID: 23601690

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Abstract

Background Detection and validation of inhibitors (antibodies) to hemophilia treatment products are important for clinical care, evaluation of product safety and assessment of population trends. Methods Centralized monitoring for factor VIII (FVIII) inhibitors was conducted for patients in the Hemophilia Inhibitor Research Study using a previously reported modified Nijmegen-Bethesda clotting assay (NBA), a chromogenic Bethesda assay (CBA) and a novel fluorescence immunoassay (FLI). Results NBA and CBA were performed on 1005 specimens and FLI on 272 specimens. CBA was negative on 880/883 specimens (99.7%) with Nijmegen-Bethesda units (NBU) < 0.5 and positive on 42/42 specimens (100%) with NBU ≥ 2.0 and 43/80 specimens (53.8%) with NBU 0.5–1.9. Among specimens with positive NBA and negative CBA, 58.1% were FLI negative, 12.9% had evidence of lupus anticoagulant, and 35.5% had non-time-dependent inhibition. CBA and FLI were positive on 72.4% and 100% of 1.0–1.9 NBU specimens and 43.1% and 50.0% of 0.5–0.9 NBU specimens. FLI detected antibodies in 98.0% of CBA-positive and 81.6% of NBA-positive specimens (P = 0.004). Among 21 new inhibitors detected by NBA, five (23.8%) with 0.7–1.3 NBU did not react in CBA or FLI. Among previously positive patients with 0.5–1.9 NBU, 7/25 (28%) were not CBA or FLI positive. FLI was positive on 36/169 NBU-negative specimens (21.3%). Conclusions FVIII specificity could not be demonstrated by CBA or FLI for 26% of inhibitors of 0.5–1.9 NBU; such results must be interpreted with caution. Low titer inhibitors detected in clot-based assays should always be repeated, with consideration given to evaluating their reactivity with FVIII using more specific assays.

fluorescence immunoassay

factor VIII inhibitor

chromogenic factor VIII assay

Nijmegen-Bethesda assay

Details

Title: Subtitle: Comparison of Clot-based, Chromogenic, and Fluorescence Assays for Measurement of Factor VIII Inhibitors in the U.S. Hemophilia Inhibitor Research Study
Creators: Connie H Miller - Division of Blood Disorders, National Center on Birth Defects and Developmental Disabilities, Centers for Disease Control and Prevention, Atlanta, Georgia
Anne S Rice - Division of Blood Disorders, National Center on Birth Defects and Developmental Disabilities, Centers for Disease Control and Prevention, Atlanta, Georgia
Brian Boylan - Division of Blood Disorders, National Center on Birth Defects and Developmental Disabilities, Centers for Disease Control and Prevention, Atlanta, Georgia
Amy D Shapiro - Indiana Hemophilia and Thrombosis Center, Indianapolis, IN
Steven R Lentz - University of Iowa Carver College of Medicine, Iowa City, IA
Brian M Wicklund - Kansas City Regional Hemophilia Center, Kansas City, MO
Fiona M Kelly - Division of Blood Disorders, National Center on Birth Defects and Developmental Disabilities, Centers for Disease Control and Prevention, Atlanta, Georgia
J. Michael Soucie - Division of Blood Disorders, National Center on Birth Defects and Developmental Disabilities, Centers for Disease Control and Prevention, Atlanta, Georgia
Hemophilia Inhibitor Research Study Investigators
Resource Type: Journal article
Publication Details: Journal of thrombosis and haemostasis, Vol.11(7), pp.1300-1309
DOI: 10.1111/jth.12259
PMID: 23601690
PMCID: PMC4477744
NLM abbreviation: J Thromb Haemost
ISSN: 1538-7933
eISSN: 1538-7836
Grant note: DOI: 10.13039/100014476, name: Pfizer Pharmaceuticals; name: Baxter Healthcare; name: CDC Foundation
Language: English
Date published: 07/2013
Academic Unit: Hematology, Oncology, and Blood & Marrow Transplantation; Internal Medicine
Record Identifier: 9984094338002771

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