title: Analysis of a high-throughput HLA antibody screening assay for use with platelet donors.
abstract: BACKGROUND
  Passive infusion of HLA antibodies has been implicated in transfusion reactions. A rapid, inexpensive method of screening blood donors for HLA antibodies might reduce the incidence of reactions. A high-throughput microbead-flow analyzer HLA antibody detection technique was compared with an enzyme-linked immunosorbent assay (ELISA) method.
  
  
  METHODS
  Ninety-six apheresis platelet (PLT) donors were tested for antibodies to Class I and II HLA antigens with mixed-antigen microbead-flow analyzer and ELISAs. For both assays, samples reactive in the mixed-antigen assay were tested with a panel-reactive antibody (PRA) assay. Samples reactive in both the mixed-antigen and the PRA assays were considered positive.
  
  
  RESULTS
  In the mixed-antigen microbead assay, 46 (48%) samples were reactive to Class I antigens and 20 (21%) to Class II. Further testing in the microbead PRA assay revealed that 34 (35%) had antibodies to Class I antigens, 18 (19%) to Class II, and 42 (44%) to either Class I or Class II. Class I antibodies were present in 56 percent of females and 36 percent of males. In the mixed-antigen ELISA, 4 samples were reactive with Class I antigens, 4 with Class II antigens, and 5 with Class I or Class II. All 5 reactive samples were also reactive in the ELISA PRA assay and were from females.
  
  
  CONCLUSIONS
  The microbead assay was more sensitive than the ELISA and detected antibodies in a large proportion of donors. Samples reactive in the mixed-antigen microbead assay should be confirmed by a second assay before concluding that antibodies are present.
authors.affiliation: Department of Transfusion Medicine, Clinical Center, NIH, Bethesda, MD 20892, USA.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Fadeyi
authors.forename: Sharon
authors.initials: S
authors.lastname: Adams
authors.forename: Brett
authors.initials: B
authors.lastname: Peterson
authors.forename: Julia
authors.initials: J
authors.lastname: Hackett
authors.forename: Phyllis
authors.initials: P
authors.lastname: Byrne
authors.forename: Harvey G
authors.initials: HG
authors.lastname: Klein
authors.forename: Francesco M
authors.initials: FM
authors.lastname: Marincola
authors.forename: Susan F
authors.initials: SF
authors.lastname: Leitman
authors.forename: David F
authors.initials: DF
authors.lastname: Stroncek
mesh_terms: D000906:Antibodies / Q000276:immunology*; D001782:Blood Donors; D001792:Blood Platelets / Q000276:immunology*; D005260:Female; D006680:HLA Antigens / Q000097:blood* / Q000276:immunology*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008297:Male; D008863:Microspheres; D017713:Platelet Transfusion
chemical_list: D000906:Antibodies; D006680:HLA Antigens