Verification of the Automated ELISA Assay for Hepcidin-25 in Human Serum

Authors

  • Tara Rolić Department of Chemistry, Biochemistry and Clinical Chemistry, Faculty of Medicine, University of Osijek, Osijek, Croatia; Institute of Clinical Laboratory Diagnostics, Osijek University Hospital, Osijek, Croatia Author https://orcid.org/0000-0003-1731-6645
  • Sanja Mandić Department of Chemistry, Biochemistry and Clinical Chemistry, Faculty of Medicine, University of Osijek, Osijek, Croatia; Institute of Clinical Laboratory Diagnostics, Osijek University Hospital, Osijek, Croatia Author https://orcid.org/0000-0003-3921-8181
  • Iva Lukić Department of Chemistry, Biochemistry and Clinical Chemistry, Faculty of Medicine, University of Osijek, Osijek, Croatia; Institute of Clinical Laboratory Diagnostics, Osijek University Hospital, Osijek, Croatia Author
  • Vesna Horvat Department of Chemistry, Biochemistry and Clinical Chemistry, Faculty of Medicine, University of Osijek, Osijek, Croatia; Medical Biochemistry Laboratory, Health Center Osijek-Baranja County, Osijek, Croatia Author https://orcid.org/0000-0001-9554-4278
  • Ines Banjari Department of Food and Nutrition Research, Faculty of Food Technology Osijek, Osijek, Croatia Author

DOI:

https://doi.org/10.26332/seemedj.v7i1.266

Keywords:

enzyme-linked immunosorbent assay, hepcidins, homeostasis, iron

Abstract

Introduction: Hepcidin-25, the bioactive form of hepcidin, is the master protein in regulating iron homeostasis. Serum concentrations, measured by different methods, are often incomparable and complicate results interpretation.

Materials and Methods: The aim was to verify the first fully automated enzyme-linked immunosorbent assay (ELISA) method, using the DRG Hybrid XL analyzer (DRG Instruments, Marburg, Germany) standardized against the mass spectrometry method. Intra- (CVi) and inter-assay (CVg) precision and bias were performed using commercially available controls with low (C1) and high (C2) concentrations. The reference interval was verified by analyzing serum samples of 20 healthy males.

Results: CVi = 9.1% (C1), 4.5% (C2); CVg = 8.9% (C1), 5.6% (C2); calculated bias was 33% for C1 and 20% for C2, respectively.

Conclusion: Verification of the fully automated ELISA method for hepcidin-25 in serum on the DRG Hybrid XL analyzer met the analytical acceptance criteria.

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Published

2023-04-30

Issue

Vol. 7 No. 1 (2023)

Section

Articles

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Copyright (c) 2025 Southeastern European Medical Journal

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