25-hydroxyvitamin D; LC-MS/MS; analytical performance specifications; immunoassays; measurement uncertainty; standardization; Vitamin D; Humans; Immunoassay/standards; Immunoassay/methods; Chromatography, Liquid/methods; Chromatography, Liquid/standards; Uncertainty; Liquid Chromatography-Mass Spectrometry; Tandem Mass Spectrometry/methods; Tandem Mass Spectrometry/standards; Vitamin D/blood; Vitamin D/analogs & derivatives; Chromatography, Liquid; Immunoassay; Tandem Mass Spectrometry; Clinical Biochemistry; Biochemistry (medical)
Abstract :
[en] [en] OBJECTIVES: Accurate 25-hydroxyvitamin D (25-(OH)D) assays are essential for defining vitamin D status and ensuring appropriate clinical decisions. Standardization efforts, including the Vitamin D Standardization Program (VDSP), aim to minimize assay variability. This study evaluates the measurement uncertainty (MU) of various 25-(OH)D assays and their ability to detect physiologically relevant changes over time.
METHODS: Seventeen pooled and eight single-donor serum samples were analyzed using two LC-MS/MS methods and 13 immunoassays, each applied in two independent laboratories. Imprecision, bias, and MU were assessed relative to the University of Ghent's reference measurement procedure (RMP). Results were compared against analytical performance specifications (APS) from VDSP, JCTLM-TF-RMSI, and IFCC C-BM based on physiological 25-(OH)D variation. A graphical approach was introduced to visualize MU in relation to clinical relevance.
RESULTS: LC-MS/MS methods consistently met all APS criteria. Several immunoassays also achieved acceptable MU, although significant bias or inter-laboratory variability was observed for some of them. Slightly more than half of the assays met the desirable Joint Committee for Traceability in Laboratory Medicine Task Force on Reference Measurement System Implementation (JCTLM TF-RMSI) MU threshold (≤10 %), while four exceeded the minimum acceptable limit (≤15 %). The IFCC C-BM physiological approach identified a similar subset of assays. The graphical representation effectively illustrated method reliability across the tested concentration range.
CONCLUSIONS: Measurement uncertainty remains a major challenge for 25-(OH)D assays. The integration of MU-based APS and graphical visualization provides a comprehensive framework for evaluating assay performance. These findings highlight the importance of selecting assays capable of reliably detecting clinically meaningful changes in vitamin D status.
Disciplines :
Laboratory medicine & medical technology
Author, co-author :
Cavalier, Etienne ; Université de Liège - ULiège > Département de pharmacie > Chimie médicale
Bhattoa, Harjit P ; Department of Laboratory Medicine, Faculty of Medicine, University of Debrecen, Debrecen, Hungary
Heijboer, Annemieke C; Department of Laboratory Medicine, Endocrine Laboratory, Amsterdam UMC, Location Vrije Universiteit Amsterdam and Location University of Amsterdam, Amsterdam, The Netherlands
Lombardi, Giovanni ; Laboratory of Experimental Biochemistry, IRCCS Ospedale Galeazzi-Sant'Ambrogio, Milan, Italy
Delatour, Vincent; Laboratoire National de Métrologie et D'Essais (LNE), Paris, France
Van Uytfanghe, Katleen; Department of Bioanalysis, Faculty of Pharmaceutical Sciences, Ref4U-Laboratory of Toxicology, Ghent University, Ghent, Belgium
Makris, Konstantinos ; Laboratory for the Research of Musculoskeletal System "Th. Garofalidis", School of Medicine, National and Kapodistrian, University of Athens, Athens, Greece
Jørgensen, Niklas Rye; Department of Clinical Biochemistry, Copenhagen University Hospital-Rigshospitalet, Copenhagen, Denmark ; Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
Herrmann, Markus; Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University of Graz, Graz, Austria
Pikner, Richard; Department of Clinical Biochemistry and Bone metabolism, Klatovska Hospital, Klatovy, Czech Republic ; Department of Clinical Biochemistry and Haematology, Faculty of Medicine Pilsen, Charles University Prague, Pilsen, Czech Republic ; Faculty of Health Care Studies, University of West Bohemia, Pilsen, Czech Republic
LUKAS, Pierre ; Centre Hospitalier Universitaire de Liège - CHU > > Service de chimie clinique
Vasikaran, Samuel D; PathWest Laboratory Medicine, Fiona Stanley Hospital, Murdoch, WA, Australia
Fraser, Callum G ; Population Health and Genomics, Ninewells Hospital and Medical School, University of Dundee, Dundee, UK
We would like to thank the colleagues who kindly accepted to measure the samples: Inge De Cuyper and Dieter Desmet (AZ Delta), Marie-Louise Schleck (H\u00F4pital de la Citadelle), Lucas Dewalque (CHC Mont-L\u00E9gia), Emilie Catry (CHU UCL Mont-Godinne), Fleur Wolf, and Fr\u00E9d\u00E9ric Cotton (LHUB Bruxelles).
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