HLA antibody detection and characterization by solid phase immunoassays: methods and pitfalls.
Solid phase immunoassays for the detection and characterization of HLA-specific antibodies provide greatly increased sensitivity, specificity, and time and reagent efficiency, compared to the traditionally used cell-based methods. Testing is performed using commercially available test kits. The assays are of two general types: enzyme-linked immunosorbent assays and multianalyte bead. The types vary in both sensitivity and equipment requirements.While these assays afford great improvement over the cell-based assays, they can be confounded by interference from substances within the serum that result in high background reactivity. The high sensitivity of the assays also makes them more susceptible to environmental factors and operator variability. The user must be aware of the capabilities of the various formats, the factors that can affect test results, and lot to lot variability of any single product. Knowledge of the characteristics of each product and thorough and accurate analysis of the results are essential to the utility of these assays.
Immunogenetics Laboratory, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA. aaz@jhmi.edu
Andrea A
AA
Zachary
Renato M
RM
Vega
Donna P
DP
Lucas
Mary S
MS
Leffell
D000906:Antibodies / Q000032:analysis* / Q000276:immunology*; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D006680:HLA Antigens / Q000276:immunology*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D012984:Software
D000906:Antibodies; D006680:HLA Antigens