Analysis of a high-throughput HLA antibody screening assay for use with platelet donors.
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.
Fadeyi
Emmanuel
E
Department of Transfusion Medicine, Clinical Center, NIH, Bethesda, MD 20892, USA.
Adams
Sharon
S
Peterson
Brett
B
Hackett
Julia
J
Byrne
Phyllis
P
Klein
Harvey G
HG
Marincola
Francesco M
FM
Leitman
Susan F
SF
Stroncek
David F
DF
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
D000906:Antibodies; D006680:HLA Antigens