title: How can we reduce costs of solid-phase multiplex-bead assays used to determine anti-HLA antibodies?
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.
authors.affiliation: Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: E G
authors.initials: EG
authors.lastname: Kamburova
authors.affiliation: Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: B W
authors.initials: BW
authors.lastname: Wisse
authors.affiliation: Laboratory Medicine, Lab. Medical Immunology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: I
authors.initials: I
authors.lastname: Joosten
authors.affiliation: Laboratory Medicine, Lab. Medical Immunology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: W A
authors.initials: WA
authors.lastname: Allebes
authors.affiliation: Laboratory Medicine, Lab. Medical Immunology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: A
authors.initials: A
authors.lastname: van der Meer
authors.affiliation: Department of Nephrology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: L B
authors.initials: LB
authors.lastname: Hilbrands
authors.affiliation: Department of Nephrology, Radboud University Medical Center, Nijmegen, The Netherlands.
authors.forename: M C
authors.initials: MC
authors.lastname: Baas
authors.affiliation: Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: E
authors.initials: E
authors.lastname: Spierings
authors.affiliation: Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: C E
authors.initials: CE
authors.lastname: Hack
authors.affiliation: Department of Nephrology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: F E
authors.initials: FE
authors.lastname: van Reekum
authors.affiliation: Department of Nephrology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: A D
authors.initials: AD
authors.lastname: van Zuilen
authors.affiliation: Department of Nephrology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: M
authors.initials: M
authors.lastname: Verhaar
authors.affiliation: Julius Center for Health Sciences and Primary Care, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: M L
authors.initials: ML
authors.lastname: Bots
authors.affiliation: Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: A C A D
authors.initials: AC
authors.lastname: Drop
authors.affiliation: Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: L
authors.initials: L
authors.lastname: Plaisier
authors.affiliation: Department of Nephrology, University of Groningen, University Medical Center, Groningen, The Netherlands.
authors.forename: M A J
authors.initials: MA
authors.lastname: Seelen
authors.affiliation: Department of Nephrology, University of Groningen, University Medical Center, Groningen, The Netherlands.
authors.forename: J S F
authors.initials: JS
authors.lastname: Sanders
authors.affiliation: Department of Laboratory Medicine, University of Groningen, University Medical Center, Groningen, The Netherlands.
authors.forename: B G
authors.initials: BG
authors.lastname: Hepkema
authors.affiliation: Department of Laboratory Medicine, University of Groningen, University Medical Center, Groningen, The Netherlands.
authors.forename: A J
authors.initials: AJ
authors.lastname: Lambeck
authors.affiliation: Department of Laboratory Medicine, University of Groningen, University Medical Center, Groningen, The Netherlands.
authors.forename: L B
authors.initials: LB
authors.lastname: Bungener
authors.affiliation: Department of Laboratory Medicine, University of Groningen, University Medical Center, Groningen, The Netherlands.
authors.forename: C
authors.initials: C
authors.lastname: Roozendaal
authors.affiliation: Department of Transplantation Immunology, Tissue Typing Laboratory, Maastricht University Medical Centre, Maastricht, The Netherlands.
authors.forename: M G J
authors.initials: MG
authors.lastname: Tilanus
authors.affiliation: Department of Transplantation Immunology, Tissue Typing Laboratory, Maastricht University Medical Centre, Maastricht, The Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: Vanderlocht
authors.affiliation: Department of Transplantation Immunology, Tissue Typing Laboratory, Maastricht University Medical Centre, Maastricht, The Netherlands.
authors.forename: C E
authors.initials: CE
authors.lastname: Voorter
authors.affiliation: Department of Transplantation Immunology, Tissue Typing Laboratory, Maastricht University Medical Centre, Maastricht, The Netherlands.
authors.forename: L
authors.initials: L
authors.lastname: Wieten
authors.affiliation: Department of Internal Medicine, Division of Nephrology, Maastricht University Medical Centre, Maastricht, The Netherlands.
authors.forename: E M
authors.initials: EM
authors.lastname: van Duijnhoven
authors.affiliation: Department of Internal Medicine, Division of Nephrology, Maastricht University Medical Centre, Maastricht, The Netherlands.
authors.forename: M
authors.initials: M
authors.lastname: Gelens
authors.affiliation: Department of Internal Medicine, Division of Nephrology, Maastricht University Medical Centre, Maastricht, The Netherlands.
authors.forename: M H L
authors.initials: MH
authors.lastname: Christiaans
authors.affiliation: Department of Nephrology, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: F J
authors.initials: FJ
authors.lastname: van Ittersum
authors.affiliation: Department of Nephrology, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: A
authors.initials: A
authors.lastname: Nurmohamed
authors.affiliation: Department of Immunogenetics, Sanquin, Amsterdam, The Netherlands.
authors.forename: N M
authors.initials: NM
authors.lastname: Lardy
authors.affiliation: Department of Immunogenetics, Sanquin, Amsterdam, The Netherlands.
authors.forename: W
authors.initials: W
authors.lastname: Swelsen
authors.affiliation: Renal Transplant Unit, Department of Internal Medicine, Academic Medical Centre, Amsterdam, The Netherlands.
authors.forename: K A
authors.initials: KA
authors.lastname: van der Pant
authors.affiliation: Renal Transplant Unit, Department of Internal Medicine, Academic Medical Centre, Amsterdam, The Netherlands.
authors.forename: N C
authors.initials: NC
authors.lastname: van der Weerd
authors.affiliation: Renal Transplant Unit, Department of Internal Medicine, Academic Medical Centre, Amsterdam, The Netherlands.
authors.forename: I J M
authors.initials: IJ
authors.lastname: Ten Berge
authors.affiliation: Renal Transplant Unit, Department of Internal Medicine, Academic Medical Centre, Amsterdam, The Netherlands.
authors.forename: F J
authors.initials: FJ
authors.lastname: Bemelman
authors.affiliation: Dutch Organ Transplant Registry (NOTR), Dutch Transplant Foundation (NTS), Leiden, The Netherlands.
authors.forename: A
authors.initials: A
authors.lastname: Hoitsma
authors.affiliation: Department of Nephrology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: P J M
authors.initials: PJ
authors.lastname: van der Boog
authors.affiliation: Department of Nephrology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: J W
authors.initials: JW
authors.lastname: de Fijter
authors.affiliation: Department of Internal Medicine, Erasmus MC, Rotterdam, The Netherlands.|Department of Nephrology, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: M G H
authors.initials: MG
authors.lastname: Betjes
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: S
authors.initials: S
authors.lastname: Heidt
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: D L
authors.initials: DL
authors.lastname: Roelen
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: F H
authors.initials: FH
authors.lastname: Claas
authors.affiliation: Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: H G
authors.initials: HG
authors.lastname: Otten
mesh_terms: D000483:Alleles; D057205:Automation, Laboratory / Q000191:economics* / Q000592:standards; D006680:HLA Antigens / Q000097:blood / Q000276:immunology*; D006650:Histocompatibility Testing; D006801:Humans; D007106:Immune Sera / Q000737:chemistry; D007118:Immunoassay / Q000191:economics* / Q000592:standards; D007518:Isoantibodies / Q000097:blood*; D016030:Kidney Transplantation; D015588:Observer Variation; D011933:Reagent Kits, Diagnostic / Q000191:economics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D006680:HLA Antigens; D007106:Immune Sera; D007518:Isoantibodies; D011933:Reagent Kits, Diagnostic
keywords: human leukocyte antigen antibodies; luminex; single antigen; solid-phase multiplex-bead assays