Application of the particle gel agglutination assay in the typing of single human leucocyte antigens

Abstract

We describe a simple and rapid particle gel agglutination assay (PaGIA) for typing of the human leucocyte antigens (HLA) HLA-A2, HLA-B7 and HLA-B27. Superparamagnetic streptavidin particles were coated with biotinylated monoclonal antibodies (MoAbs) to HLA-A2, HLA-B7 and HLA-B27. Anticoagulated whole blood samples from healthy blood donors (n = 118) with known HLA patterns were incubated with MoAb-coated particles, transferred into a standard ID-gel card, and subsequently centrifuged. Samples were evaluated macroscopically, with antigen-positive samples resulting in a visible agglutination reaction. A clear distinction could be made between all positive and negative samples tested. Fifty-seven samples were found to be positive for HLA-A2 (48%), 26 samples for HLA-B7 (22%) and 5 samples for HLA-B27 (4%).

Rapid typing of the human neutrophil antigen 1a by the particle gel agglutination assay

“…The described HNA-1a typing method requires no additional laboratory equipment, has been shown to be feasible for the typing of other antigens (e.g. HLA-A2, HLA-B27, and HLA-B7; HPA-1a; and HNA-1a) (8)(9)(10)(11), and may be expand for the typing of a large variety other antigens if specific MoAbs are available. Figure 1 Results of human neutrophil antigen (HNA)-1a phenotyping of four different blood samples by the particle agglutination assay using anti-HNA-1a-coated superparamagnetic particles.…”

“…The described HNA‐1a typing method requires no additional laboratory equipment, has been shown to be feasible for the typing of other antigens (e.g. HLA‐A2, HLA‐B27, and HLA‐B7; HPA‐1a; and HNA‐1a) (8–11), and may be expand for the typing of a large variety other antigens if specific MoAbs are available.…”