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Southeastern European Medical Journal : SEEMEDJ, Vol. 7 No. 1, 2023.

Original scientific paper

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

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

Tara Rolić orcid id orcid.org/0000-0003-1731-6645 ; 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
Sanja Mandić orcid id orcid.org/0000-0003-3921-8181 ; 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
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
Vesna Horvat orcid id orcid.org/0000-0001-9554-4278 ; 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
Ines Banjari orcid id orcid.org/0000-0002-8680-5007 ; Department of Food and Nutrition Research, Faculty of Food Technology Osijek, Osijek, Croatia

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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.

Keywords

enzyme-linked immunosorbent assay; hepcidins; homeostasis; iron

Hrčak ID:

301084

URI

https://hrcak.srce.hr/301084

Publication date:

30.4.2023.

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