Optimizing hepcidin measurement with a proficiency test framework and standardization improvement

• Published in: Clinical chemistry and laboratory medicine Vol. 59; no. 2; pp. 315 - 323
• Main Authors: Aune, Ellis T., Diepeveen, Laura E., Laarakkers, Coby M., Klaver, Siem, Armitage, Andrew E., Bansal, Sukhvinder, Chen, Michael, Fillet, Marianne, Han, Huiling, Herkert, Matthias, Itkonen, Outi, van de Kerkhof, Daan, Krygier, Aleksandra, Lefebvre, Thibaud, Neyer, Peter, Rieke, Markus, Tomosugi, Naohisa, Weykamp, Cas W., Swinkels, Dorine W.
• Format: Journal Article Web Resource
• Language: English
• Published: Germany De Gruyter 23.02.2021; Walter De Gruyter & Company; Walter de Gruyter
• Subjects: Accreditation; Blood Specimen Collection; Calibration; Chromatography, High Pressure Liquid; Enzyme-Linked Immunosorbent Assay; external quality assurance; Hepcidin; Hepcidins - blood; Homogeneity; Human health sciences; Humans; iron metabolism; Laboratories - standards; Linearity; Optimization; Pharmacie, pharmacologie & toxicologie; Pharmacy, pharmacology & toxicology; proficiency testing; Quality assurance; Quality Assurance, Health Care - standards; Quality Control; Reference Standards; Sciences de la santé humaine; secondary reference material; Standardization; Tandem Mass Spectrometry; iron metabolism; standardization; hepcidin; proficiency testing; secondary reference material; external quality assurance
• ISSN: 1434-6621; 1437-4331; 1437-4331
• DOI: 10.1515/cclm-2020-0928

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. 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. 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. 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.