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Validation of a competitive ELISA assay for the quantification of human serum hepcidin

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.

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Hepcidin -25 (bioactive) HS Dahlfors G1, Stål P2, Hansson EC1, Bàràny P3, Sisowath C1, Onelöv L1, Nelson D4, Eggertsen G1, Marmur J5, Beshara S1.

Author Information

Scand J Clin Lab Invest. 2015;75(8):652-8. Epub 2015 Aug 12.

Link: https://www.ncbi.nlm.nih.gov/pubmed/26264426

by Oleg Vishnevski | Aug 12, 2015

 
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