학술논문
How can we reduce costs of solid-phase multiplex-bead assays used to determine anti- HLA antibodies?
Document Type
Article
Author
Kamburova, E. G.; Wisse, B. W.; Joosten, I.; Allebes, W. A.; van der Meer, A.; Hilbrands, L. B.; Baas, M. C.; Spierings, E.; Hack, C. E.; van Reekum, F. E.; van Zuilen, A. D.; Verhaar, M.; Bots, M. L.; Drop, A. C. A. D.; Plaisier, L.; Seelen, M. A. J.; Sanders, J. S. F.; Hepkema, B. G.; Lambeck, A. J.; Bungener, L. B.
Source
Subject
*HLA histocompatibility antigens
*IMMUNOGLOBULINS
*SOLID-phase analysis
*BIOLOGICAL assay
*BIOLOGICAL reagents
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Language
ISSN
2059-2302
Abstract
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. [ABSTRACT FROM AUTHOR]