Mass spectrometry measurement of plasma hepcidin for the prediction of iron overload
-
Annarosa Floreani
,
Filippo Navaglia
Abstract
Background: Hepcidin has emerged as the primary regulator of iron homeostasis. Previous studies on assessing urinary hepcidin are limited. We developed a method for quantifying hepcidin-25 (Hep-25) in plasma using surface-enhanced laser-desorption-ionization time-of-flight mass spectrometry (SELDI-TOF/MS) and a 25-AA peptide as reference standard. The aims of the study were 1) to assess the performance of this method in different conditions of iron metabolism disorders; 2) to assess the diagnostic validity of non-invasive serum biomarkers in the identification of iron overload.
Methods: Validation of the method was performed in 10 patients with type I hemochromatosis (HE) and in 177 subjects previously enrolled in a general population epidemiological study. Among the latter group, 17 had non-alcoholic fatty liver disease, 10 had chronic hepatitis C, and 150 subjects had normal ultrasound, normal liver function tests (LFTs), an alcohol intake <20 g ethanol/day and were negative for the C282Y mutation. The following biomarkers were assayed in each case: plasma Hep-25, C282Y and H63D mutations of the HFE gene; serum iron, ferritin (SF), transferrin saturation, transaminases, γ-glutamyltransferase (GGT), glucose, insulin, total cholesterol, high-density lipoprotein (HDL)-cholesterol, low-density lipoprotein (LDL)-cholesterol and triglycerides.
Results: Plasma Hep-25 concentrations were higher in HCV+ patients (26.3±7.2 nmol/L) than in controls, and correlated positively with SF (p<0.001). H63D heterozygous subjects revealed a pattern of iron overload that was significantly higher than H63D wild type subjects. Analyzing the data with the Biomarker Pattern 5.0.2. software to identify the most significant biomarkers for discriminating between HE cases and controls allowed us to produce an algorithm with four terminal nodes, which included glucose >4.8 mmol/L and Hep-25/SF ratio ≤6.6 as the main splitters. These variables enabled the correct diagnosis of HE with 100% sensitivity, 93% specificity and an area under the receiver operating characteristic (ROC) curve of 0.993.
Conclusions: Our plasma Hep-25 mass spectrometry method yields measurements that reflect pathological and genetic influences; simple non-invasive biomarkers (Hep-25/SF ratio and glucose) can predict the presence of HE.
©2011 by Walter de Gruyter Berlin New York
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Articles in the same Issue
- Editorial
- Hepcidin: the main regulator of iron homeostasis
- Reviews
- Global trends in critical values practices and their harmonization
- Ischemia modified albumin changes – review and clinical implications
- Genetics and Molecular Diagnostics
- Decreased serum brain-derived neurotrophic factor (BDNF) is associated with post-stroke depression but not with BDNF gene Val66Met polymorphism
- A novel liquidchip platform for simultaneous detection of 70 alleles of DNA somatic mutations on EGFR, KRAS, BRAF and PIK3CA from formalin-fixed and paraffin-embedded slides containing tumor tissue
- General Clinical Chemistry and Laboratory Medicine
- Mass spectrometry measurement of plasma hepcidin for the prediction of iron overload
- Serum hepcidin-25 in comparison to biochemical markers and hematological indices for the differentiation of iron-restricted erythropoiesis
- The quality of the extra-analytical phase of laboratory practice in some developing European countries and Mexico – a multicentric study
- Impact of cryopreservation on serum concentration of matrix metalloproteinases (MMP)-7, TIMP-1, vascular growth factors (VEGF) and VEGF-R2 in Biobank samples
- Determinants of circulating asymmetric and symmetric dimethylarginines in patients evaluated for acute dyspnea
- Urinary stone composition in pediatric patients: a retrospective study of 205 cases
- Electrolyte-balanced heparin in blood gas syringes can introduce a significant bias in the measurement of positively charged electrolytes
- Full-scan mass spectral evidence for 3-epi-25-hydroxyvitamin D3 in serum of infants and adults
- Association of nucleated red blood cells in blood and arterial oxygen partial tension
- Validation and Outcome Studies
- Cystatin C is a reliable marker for estimation of glomerular filtration rate in renal transplantation: validation of a new turbidimetric assay using monospecific sheep antibodies
- Analytical validation of the Liaison Calcitonin_II-Gen (DiaSorin)
- A new tool in the field of in-vitro diagnosis of allergy: preliminary results in the comparison of ImmunoCAP© 250 with the ImmunoCAP© ISAC
- Rapid microwave digestion and microplate reading format method for urinary iodine determination
- Reference Values and Biological Variations
- Serum creatinine concentrations in male and female elite swimmers. Correlation with body mass index and evaluation of estimated glomerular filtration rate
- Biological variation of free β chorionic gonadotropin and pregnancy-associated plasma protein A in first trimester pregnancies
- Non-infectious conditions and gestational age influence C-reactive protein values in newborns during the first 3 days of life
- Cancer Diagnostics
- Interpretation of increments in serial tumour biomarker concentrations depends on the distance of the baseline concentration from the cut-off
- Performance of nuclear matrix protein 22 urine marker and voided urine cytology in the detection of urinary bladder tumors
- High-performance metabolic marker assessment in breast cancer tissue by mass spectrometry
- BRAF analysis by fine needle aspiration biopsy of thyroid nodules improves preoperative identification of papillary thyroid carcinoma and represents a prognostic factor. A mono-institutional experience
- Letters to the Editor
- Stat laboratory timeliness management according to clinician needs
- Stability of serum samples and hemolysis interference on the high sensitivity troponin T assay
- Falsely diminished results for insulin concentrations measured with the IMMULITE® 2000
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