Solid-phase multiplex-bead assays are widely used in transplantation to detect anti-human leukocyte antigen (HLA) antibodies. These assays enable high resolution detection of low levels of HLA antibodies. However, multiplex-bead assays are costly and yield variable measurements that limit the comparison of results between laboratories. In the context of a Dutch national Consortium study we aimed to determine the inter-assay and inter-machine variability of multiplex-bead assays, and we assessed how to reduce the assay reagents costs. Fifteen sera containing a variety of HLA antibodies were used yielding in total 7092 median fluorescence intensities (MFI) values. The inter-assay and inter-machine mean absolute relative differences (MARD) of the screening assay were 12% and 13%, respectively. The single antigen bead (SAB) inter-assay MARD was comparable, but showed a higher lot-to-lot variability. Reduction of screening assay reagents to 50% or 40% of manufacturers' recommendations resulted in MFI values comparable to 100% of the reagents, with an MARD of 12% or 14%, respectively. The MARD of the 50% and 40% SAB assay reagent reductions were 11% and 22%, respectively. From this study, we conclude that the reagents can be reliably reduced at least to 50% of manufacturers' recommendations with virtually no differences in HLA antibody assignments.

Additional Metadata
Keywords human leukocyte antigen antibodies, luminex, single antigen, solid-phase multiplex-bead assays
Persistent URL dx.doi.org/10.1111/tan.12860, hdl.handle.net/1765/97439
Journal HLA
Citation
Kamburova, E.G, Wisse, B.W, Joosten, I, Allebes, W.A, van der Meer, A, Hilbrands, L.B, … Otten, H.G. (2016). How can we reduce costs of solid-phase multiplex-bead assays used to determine anti-HLA antibodies?. HLA, 88(3), 110–119. doi:10.1111/tan.12860