Ellis T Aune1,2, Laura E Diepeveen1,2, Coby M Laarakkers1,2, Siem Klaver1,2, Andrew E Armitage3, Sukhvinder Bansal4, Michael Chen5,6, Marianne Fillet7, Huiling Han8, Matthias Herkert9, Outi Itkonen10, Daan van de Kerkhof11, Aleksandra Krygier12, Thibaud Lefebvre13, Peter Neyer14, Markus Rieke15, Naohisa Tomosugi16, Cas W Weykamp17,18, Dorine W Swinkels1,2,19. 1. Department of Laboratory Medicine, Radboud University Medical Center, Nijmegen, The Netherlands. 2. Hepcidinanalysis.com, Nijmegen, The Netherlands. 3. MRC Human Immunology Unit, MRC Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, UK. 4. Department of Pharmacy, School of Cancer and Pharmaceutical Science, King's College London, London, UK. 5. Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, Canada. 6. Division of Medical Sciences, University of Victoria, Victoria, Canada. 7. Laboratory for the Analysis of Medicines, CIRM, University of Liège, Liège, Belgium. 8. Intrinsic Life Sciences, La Jolla, USA. 9. DRG Instruments, Marburg, Germany. 10. Laboratory Division HUSLAB, Helsinki University Central Hospital, Helsinki, Finland. 11. Algemeen Klinisch Laboratorium, Catharina Ziekenhuis, Eindhoven, The Netherlands. 12. Department of Endocrinology, Metabolism and Internal Medicine, Poznan University of Medical Sciences, Poznan, Poland. 13. French Center of Porphyria, INSERM UMR1149, Labex GR-Ex, Louis Mourier Hospital, APHP.Nord-Université de Paris, Paris, France. 14. Institute of Laboratory Medicine, Kantonsspital Aarau, Aarau, Switzerland. 15. IPH GMBH, Alfeld (Leine), Germany. 16. Division of Systems Bioscience for Drug Discovery, Medical Research Institute, Kanazawa Medical University, Kahoku, Japan. 17. Department of Clinical Chemistry, Queen Beatrix Hospital, Winterswijk, The Netherlands. 18. SKML, Nijmegen, The Netherlands. 19. Department of Laboratory Medicine, Translational Metabolic Laboratory (830), Radboud University Medical Center, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands.
Abstract
Objectives: Hepcidin measurement advances insights in pathophysiology, diagnosis, and treatment of iron disorders, but requires analytically sound and standardized measurement procedures (MPs). Recent development of a two-level secondary reference material (sRM) for hepcidin assays allows worldwide standardization. However, no proficiency testing (PT) schemes to ensure external quality assurance (EQA) exist and the absence of a high calibrator in the sRM set precludes optimal standardization. Methods: We developed a pilot PT together with the Dutch EQA organization Stichting Kwaliteitsbewaking Medische Laboratoriumdiagnostiek (SKML) that included 16 international hepcidin MPs. The design included 12 human serum samples that allowed us to evaluate accuracy, linearity, precision and standardization potential. We manufactured, value-assigned, and validated a high-level calibrator in a similar manner to the existing low- and middle-level sRM. Results: The pilot PT confirmed logistical feasibility of an annual scheme. Most MPs demonstrated linearity (R2>0.99) and precision (duplicate CV>12.2%), although the need for EQA was shown by large variability in accuracy. The high-level calibrator proved effective, reducing the inter-assay CV from 42.0% (unstandardized) to 14.0%, compared to 17.6% with the two-leveled set. The calibrator passed international homogeneity criteria and was assigned a value of 9.07±0.24 nmol/L. Conclusions: We established a framework for future PT to enable laboratory accreditation, which is essential to ensure quality of hepcidin measurement and its use in patient care. Additionally, we showed optimized standardization is possible by extending the current sRM with a third high calibrator, although international implementation of the sRM is a prerequisite for its success.
Objectives:Hepcidin measurement advances insights in pathophysiology, diagnosis, and treatment of iron disorders, but requires analytically sound and standardized measurement procedures (MPs). Recent development of a two-level secondary reference material (sRM) for hepcidin assays allows worldwide standardization. However, no proficiency testing (PT) schemes to ensure external quality assurance (EQA) exist and the absence of a high calibrator in the sRM set precludes optimal standardization. Methods: We developed a pilot PT together with the Dutch EQA organization Stichting Kwaliteitsbewaking Medische Laboratoriumdiagnostiek (SKML) that included 16 international hepcidin MPs. The design included 12 human serum samples that allowed us to evaluate accuracy, linearity, precision and standardization potential. We manufactured, value-assigned, and validated a high-level calibrator in a similar manner to the existing low- and middle-level sRM. Results: The pilot PT confirmed logistical feasibility of an annual scheme. Most MPs demonstrated linearity (R2>0.99) and precision (duplicate CV>12.2%), although the need for EQA was shown by large variability in accuracy. The high-level calibrator proved effective, reducing the inter-assay CV from 42.0% (unstandardized) to 14.0%, compared to 17.6% with the two-leveled set. The calibrator passed international homogeneity criteria and was assigned a value of 9.07±0.24 nmol/L. Conclusions: We established a framework for future PT to enable laboratory accreditation, which is essential to ensure quality of hepcidin measurement and its use in patient care. Additionally, we showed optimized standardization is possible by extending the current sRM with a third high calibrator, although international implementation of the sRM is a prerequisite for its success.
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