Optimizing hepcidin measurement with a proficiency test framework and standardization improvement

Ellis T. Aune; Laura E. Diepeveen; Coby M. Laarakkers; Siem Klaver; Andrew E. Armitage; Sukhvinder Bansal; Michael Chen; Marianne Fillet; Huiling Han; Matthias Herkert; Outi Itkonen; Daan van de Kerkhof; Aleksandra Krygier; Thibaud Lefebvre; Peter Neyer; Markus Rieke; Naohisa Tomosugi; Cas W. Weykamp; Dorine W. Swinkels

doi:10.1515/cclm-2020-0928

Optimizing hepcidin measurement with a proficiency test framework and standardization improvement

Ellis T. Aune, Laura E. Diepeveen, Coby M. Laarakkers, Siem Klaver, Andrew E. Armitage, Sukhvinder Bansal, Michael Chen, Marianne Fillet, Huiling Han, Matthias Herkert, Outi Itkonen, Daan van de Kerkhof, Aleksandra Krygier, Thibaud Lefebvre, Peter Neyer, Markus Rieke, Naohisa Tomosugi, Cas W. Weykamp, Dorine W. Swinkels (Corresponding author)

Chemical Biology

Research output: Contribution to journal › Article › Academic › peer-review

16 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	315-323
Number of pages	9
Journal	Clinical Chemistry and Laboratory Medicine
Volume	59
Issue number	2
Early online date	2 Oct 2020
DOIs	https://doi.org/10.1515/cclm-2020-0928
Publication status	Published - Feb 2021

Funding

Research funding : AEA is funded by the UK Medical Research Council (MC_UU_12010/3); work in Oxford was supported by NIHR Oxford Biomedical Research Centre. Research at the Poznan University of Medical Sciences funded by a PRELUDIUM 12 grant from the Polish National Centre for Science (2016/23/N/NZ5/02573).

Funders	Funder number
National Institute for Health and Care Research

Keywords

external quality assurance
hepcidin
iron metabolism
proficiency testing
secondary reference material
standardization

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10.1515/cclm-2020-0928

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Aune, E. T., Diepeveen, L. E., Laarakkers, C. M., Klaver, S., Armitage, A. E., Bansal, S., Chen, M., Fillet, M., Han, H., Herkert, M., Itkonen, O., van de Kerkhof, D., Krygier, A., Lefebvre, T., Neyer, P., Rieke, M., Tomosugi, N., Weykamp, C. W., & Swinkels, D. W. (2021). Optimizing hepcidin measurement with a proficiency test framework and standardization improvement. Clinical Chemistry and Laboratory Medicine, 59(2), 315-323. https://doi.org/10.1515/cclm-2020-0928

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abstract = "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.",

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author = "Aune, {Ellis T.} and Diepeveen, {Laura E.} and Laarakkers, {Coby M.} and Siem Klaver and Armitage, {Andrew E.} and Sukhvinder Bansal and Michael Chen and Marianne Fillet and Huiling Han and Matthias Herkert and Outi Itkonen and {van de Kerkhof}, Daan and Aleksandra Krygier and Thibaud Lefebvre and Peter Neyer and Markus Rieke and Naohisa Tomosugi and Weykamp, {Cas W.} and Swinkels, {Dorine W.}",

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Aune, ET, Diepeveen, LE, Laarakkers, CM, Klaver, S, Armitage, AE, Bansal, S, Chen, M, Fillet, M, Han, H, Herkert, M, Itkonen, O, van de Kerkhof, D, Krygier, A, Lefebvre, T, Neyer, P, Rieke, M, Tomosugi, N, Weykamp, CW & Swinkels, DW 2021, 'Optimizing hepcidin measurement with a proficiency test framework and standardization improvement', Clinical Chemistry and Laboratory Medicine, vol. 59, no. 2, pp. 315-323. https://doi.org/10.1515/cclm-2020-0928

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AU - Aune, Ellis T.

AU - Diepeveen, Laura E.

AU - Laarakkers, Coby M.

AU - Klaver, Siem

AU - Armitage, Andrew E.

AU - Bansal, Sukhvinder

AU - Chen, Michael

AU - Fillet, Marianne

AU - Han, Huiling

AU - Herkert, Matthias

AU - Itkonen, Outi

AU - van de Kerkhof, Daan

AU - Krygier, Aleksandra

AU - Lefebvre, Thibaud

AU - Neyer, Peter

AU - Rieke, Markus

AU - Tomosugi, Naohisa

AU - Weykamp, Cas W.

AU - Swinkels, Dorine W.

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N2 - 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.

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Optimizing hepcidin measurement with a proficiency test framework and standardization improvement

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