Gunilla Dahlfors1, Per Stål2, Eva C Hansson1, Peter Bàràny3, Christin Sisowath1, Liselotte Onelöv1, Dick Nelson4, Gösta Eggertsen1, Joel Marmur5, Soheir Beshara1. 1. a Department of Clinical Chemistry , Karolinska University Hospital & Institution for Laboratory Medicine , Stockholm , Sweden. 2. b Division of Gastroenterology and Hepatology , Karolinska University Hospital & Department of Medicine , Stockholm , Sweden. 3. c Renal Medicine, CLINTEC, Karolinska Institute , Stockholm , Sweden. 4. d Department of Clinical Chemistry , Helsingborg, University and Regional Laboratories Region Skåne , Stockholm , Sweden. 5. e Department of Gastroenterology , Ersta Hospital , Stockholm , Sweden.
Abstract
BACKGROUND: Hepcidin-25 is a potential marker for iron disorders with a demand for accessible assays. This study aimed to evaluate a commercial competitive enzyme-linked immunosorbent assay (cELISA) for hepcidin quantitation. METHODS: Serum samples; 95 healthy subjects (HS), six patients with iron deficiency (ID), 84 patients with liver disorders (LD) and 220 hemodialysis patients (HD), were analyzed. Controls were used for imprecision, while accuracy was evaluated by quantitating hepcidin-25 with LC-MS/MS in 149 samples. Cross-reactivity for hepcidin-20 and hepcidin-22 was tested. Hepcidin-mRNA expression in 37 liver biopsies was measured. RESULTS: S-hepcidin ranged from 8-76 and 2-31 μg/L in healthy men and women. Levels in ID, LD and HD significantly differed from HS. Total coefficients of variation (CV) for controls were 24% and 22%. Within-sample CV was 10%. Despite a good correlation with LC-MS/MS (r = 0.89), the cELISA showed higher values and detected hepcidin-20 and hepcidin-22. Hepcidin-mRNA correlated well with S-hepcidin using cELISA and LC-MS/MS (r = 0.69 and 0.64). CONCLUSIONS: The correlation with LC-MS/MS is good and the examined kit can differentiate between patient groups although it is not specific for hepcidin-25. Considering ELISA's capacity to readily be set up, the investigated kit can be applied. Specific reference ranges are required.
BACKGROUND:Hepcidin-25 is a potential marker for iron disorders with a demand for accessible assays. This study aimed to evaluate a commercial competitive enzyme-linked immunosorbent assay (cELISA) for hepcidin quantitation. METHODS: Serum samples; 95 healthy subjects (HS), six patients with iron deficiency (ID), 84 patients with liver disorders (LD) and 220 hemodialysis patients (HD), were analyzed. Controls were used for imprecision, while accuracy was evaluated by quantitating hepcidin-25 with LC-MS/MS in 149 samples. Cross-reactivity for hepcidin-20 and hepcidin-22 was tested. Hepcidin-mRNA expression in 37 liver biopsies was measured. RESULTS: S-hepcidin ranged from 8-76 and 2-31 μg/L in healthy men and women. Levels in ID, LD and HD significantly differed from HS. Total coefficients of variation (CV) for controls were 24% and 22%. Within-sample CV was 10%. Despite a good correlation with LC-MS/MS (r = 0.89), the cELISA showed higher values and detected hepcidin-20 and hepcidin-22. Hepcidin-mRNA correlated well with S-hepcidin using cELISA and LC-MS/MS (r = 0.69 and 0.64). CONCLUSIONS: The correlation with LC-MS/MS is good and the examined kit can differentiate between patient groups although it is not specific for hepcidin-25. Considering ELISA's capacity to readily be set up, the investigated kit can be applied. Specific reference ranges are required.