title: Low detection rate of antibodies to non-functional epitopes on factor VIII in patients with hemophilia A and negative for inhibitors by Bethesda assay.
abstract: In patients with hemophilia A who have an inhibitor to factor (F)VIII measured by Bethesda assay, enzyme-linked immunosorbent assay (ELISA) can also be used to detect the inhibitor. In some studies non-inhibitory antibodies were also detected by ELISA in many patients who were negative by Bethesda assay. Our aim was to investigate whether there is a higher detection rate of FVIII antibodies by ELISA compared with Bethesda assay. We also compared outcomes using three different preparations of recombinant FVIII (rFVIII) to coat the microtiter plates for ELISA. Inhibitor detection by ELISA generally agreed with the Bethesda method. Only four of 26 patients with no clinical suspicion of an inhibitor and with no detectable inhibitor by Bethesda assay showed a non-inhibitory antibody by ELISA, and three of these were only weakly positive. Patients with severe hemophilia A and the intron 22 inversion (n = 21) did not show a higher incidence of non-inhibitory antibodies compared with those without that mutation. Finally, we found that the formulation of rFVIII has a small effect on ELISA performance, mainly in detection of low-level antibody. The results of the present study are in contrast to and fail to confirm previously published reports showing a higher incidence of non-inhibitory antibodies in hemophilia A.
authors.affiliation: Division of Haematology, Institute of Medical and Veterinary Science, PO Box 14, Rundle Mall, Adelaide, SA, Australia.
authors.forename: M
authors.initials: M
authors.lastname: Ling
authors.forename: E M
authors.initials: EM
authors.lastname: Duncan
authors.forename: S E
authors.initials: SE
authors.lastname: Rodgers
authors.forename: A M
authors.initials: AM
authors.lastname: Street
authors.forename: J V
authors.initials: JV
authors.lastname: Lloyd
mesh_terms: D000328:Adult; D001323:Autoantibodies / Q000097:blood / Q000276:immunology*; D002675:Child, Preschool; D007446:Chromosome Inversion; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D005169:Factor VIII / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D006467:Hemophilia A / Q000276:immunology*; D006801:Humans; D016056:Immunodominant Epitopes / Q000276:immunology; D007438:Introns; D008875:Middle Aged; D010446:Peptide Fragments / Q000032:analysis; D011994:Recombinant Proteins / Q000032:analysis
chemical_list: D001323:Autoantibodies; C451034:B-domain-deleted factor VIII; D000939:Epitopes; D016056:Immunodominant Epitopes; D010446:Peptide Fragments; D011994:Recombinant Proteins; D005169:Factor VIII