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

Tara Rolić*; Sanja Mandić; Iva Lukić; Vesna Horvat; Ines Banjari

doi:10.26332/seemedj.v7i1.266

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 http://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 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 https://orcid.org/0000-0003-0045-841X
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 https://orcid.org/0000-0001-9554-4278
Ines Banjari Department of Food and Nutrition Research, Faculty of Food Technology Osijek, Osijek, Croatia https://orcid.org/0000-0002-8680-5007

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

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.