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Provisional standardization of hepcidin assays: creating a traceability chain with a primary reference material, candidate reference method and a commutable secondary reference material

  • Laura E. Diepeveen , Coby M.M. Laarakkers , Gustavo Martos , Marta E. Pawlak , Fatih F. Uğuz , Kim E.S.A. Verberne , Rachel P.L. van Swelm , Siem Klaver , Anton F.J. de Haan , Kelly R. Pitts , Sukhvinder S. Bansal , Ioana M. Abbas , Marianne Fillet , Thibaud Lefebvre , Anneke J. Geurts-Moespot , Domenico Girelli , Annalisa Castagna , Matthias Herkert , Outi Itkonen , Gordana Olbina , Naohisa Tomosugi , Mark E. Westerman , Vincent Delatour , Cas W. Weykamp and Dorine W. Swinkels EMAIL logo
Published/Copyright: November 29, 2018
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 57 Issue 6

Abstract

Background

Hepcidin concentrations measured by various methods differ considerably, complicating interpretation. Here, a previously identified plasma-based candidate secondary reference material (csRM) was modified into a serum-based two-leveled sRM. We validated its functionality to increase the equivalence between methods for international standardization.

Methods

We applied technical procedures developed by the International Consortium for Harmonization of Clinical Laboratory Results. The sRM, consisting of lyophilized serum with cryolyoprotectant, appeared commutable among nine different measurement procedures using 16 native human serum samples in a first round robin (RR1). Harmonization potential of the sRM was simulated in RR1 and evaluated in practice in RR2 among 11 measurement procedures using three native human plasma samples. Comprehensive purity analysis of a candidate primary RM (cpRM) was performed by state of the art procedures. The sRM was value assigned with an isotope dilution mass spectrometry-based candidate reference method calibrated using the certified pRM.

Results

The inter-assay CV without harmonization was 42.1% and 52.8% in RR1 and RR2, respectively. In RR1, simulation of harmonization with sRM resulted in an inter-assay CV of 11.0%, whereas in RR2 calibration with the material resulted in an inter-assay CV of 19.1%. Both the sRM and pRM passed international homogeneity criteria and showed long-term stability. We assigned values to the low (0.95±0.11 nmol/L) and middle concentration (3.75±0.17 nmol/L) calibrators of the sRM.

Conclusions

Standardization of hepcidin is possible with our sRM, which value is assigned by a pRM. We propose the implementation of this material as an international calibrator for hepcidin.

Keywords: harmonization; hepcidin; iron metabolism; primary reference material; secondary reference material; standardization
Corresponding author: Prof. Dr. Dorine W. Swinkels, Department of Laboratory Medicine, Translational Metabolic Laboratory (830), Radboud University Medical Center, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands; and Hepcidinanalysis.com, Nijmegen, The Netherlands, Phone: +31 (0)24-3618957, Fax: +31 (0)24-3668754

Acknowledgments

We would like to recognize our friend and colleague Dr. Mark Westerman (deceased), Founder and CEO of Intrinsic LifeSciences, LLC, La Jolla, California, for his major contribution to hepcidin research and for the development of hepcidin assays and products for use in research and clinical laboratories. We thank Drs Alexander Rennings and Mirian Janssen for their help in HFE-hemochromatosis patient inclusion, Ankie Sneek for arranging blood collection, Jochem Pastoor for his help in the design of the figures, and all patients and volunteers for their participation in the study.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: We would like to acknowledge Fondazione Cariverona. The funding organization played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

  3. Employment or leadership: LD, CL, SK, RvS and DS are employees of Radboudumc, that via its Hepcidinanalysis.com initiative offers high quality hepcidin measurements to the medical, scientific and pharmaceutical community at a fee for the service basis. KRP is an employee of Corgenix Medical Corporation specializing in IVD development, manufacturing, and distribution. MW (deceased) was President and CEO of Intrinsic LifeSciences LLC. GO is an employee of Intrinsic LifeSciences LLC and has ownership interest in the company. Intrinsic LifeSciences is engaged in the commercial development of hepcidin and related diagnostic products and provides CLIA compliant hepcidin testing to research, clinical and pharmaceutical clients worldwide.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2018-0783).

Received: 2018-07-23
Accepted: 2018-10-04
Published Online: 2018-11-29
Published in Print: 2019-05-27

©2019 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2018-0783
Keywords for this article
harmonization; hepcidin; iron metabolism; primary reference material; secondary reference material; standardization

Articles in the same Issue

  1. Frontmatter
  2. Obituary
  3. Professor Howard A. Morris
  4. Editorial
  5. The silk road to total quality in Laboratory Medicine
  6. Reviews
  7. Moving average quality control: principles, practical application and future perspectives
  8. Serum α-fetoprotein in pediatric oncology: not a children’s tale
  9. Mini Review
  10. Value-based healthcare: the role of laboratory medicine
  11. Opinion Paper
  12. Advantages and limitations of total laboratory automation: a personal overview
  13. General Clinical Chemistry and Laboratory Medicine
  14. Analysis and evaluation of the external quality assessment results of quality indicators in laboratory medicine all over China from 2015 to 2018
  15. A pilot study for establishing quality indicators in molecular diagnostics according to the IFCC WG-LEPS initiative: preliminary findings in China
  16. Quality assessment of interpretative commenting and competency comparison of comment providers in China
  17. Lower creatinine concentration values and lower inter-laboratory variation among Swedish hospital laboratories in 2014 compared to 1996: results from the Equalis external quality assessment program
  18. Development of the Point-of-Care Key Evidence Tool (POCKET): a checklist for multi-dimensional evidence generation in point-of-care tests
  19. Analytical and clinical performance evaluation of two POC tests for therapeutic drug monitoring of infliximab
  20. Hematology and Coagulation
  21. Provisional standardization of hepcidin assays: creating a traceability chain with a primary reference material, candidate reference method and a commutable secondary reference material
  22. Danger of false negative (exclusion) or false positive (diagnosis) for ‘congenital thrombophilia’ in the age of anticoagulants
  23. Point-of-care haemostasis monitoring during liver transplantation is cost effective
  24. Reference Values and Biological Variations
  25. Evaluation of reference intervals of haematological and biochemical markers in an Austrian adolescent study cohort
  26. Cancer Diagnostics
  27. A novel machine learning-derived decision tree including uPA/PAI-1 for breast cancer care
  28. Cardiovascular Diseases
  29. Evaluation of analytical performances using standardized analytical protocols and comparison of clinical results of the new ADVIA BNP and NT-proBNP immunoassays for the Centaur XPT platform
  30. Infectious Diseases
  31. Improvement in detecting sepsis using leukocyte cell population data (CPD)
  32. Letters to the Editor
  33. The biological variation of plasma proenkephalin: data from a stable heart failure cohort
  34. Hemoglobin variants found in relation to HbA1c testing: high occurrence of Hb Athens-Georgia in the Northern Jutland, Denmark
  35. Eltrombopag interferes with the measurement of plasma total bilirubin in pediatric patients in an automated colorimetric method
  36. A challenging case: highly variable TSH in a mother and her two children
  37. Suppressing all test results in grossly hemolyzed samples: is this approach appropriate in every case?
  38. Prozone effect observed for heavy chain α in the serum immunofixation electrophoresis of a patient with monoclonal IgA-λ gammopathy
  39. Significant allelic dropout phenomenon of Oncomine BRCA Research Assay on Ion Torrent S5
  40. Evaluation of immature platelet fraction in patients with myelodysplastic syndromes. Association with poor prognosis factors
  41. Influence of temperature of transport of whole blood on plasma Cu, I, Mn, Se and Zn and Mg concentrations in erythrocytes
  42. Absorbent materials to collect urine can affect proteomics and metabolomic biomarker concentrations
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