Immunoassay for human serum hepcidin

Blood ◽  
2008 ◽  
Vol 112 (10) ◽  
pp. 4292-4297 ◽  
Author(s):  
Tomas Ganz ◽  
Gordana Olbina ◽  
Domenico Girelli ◽  
Elizabeta Nemeth ◽  
Mark Westerman
Keyword(s):  
Healthy Volunteers ◽  
Immunosorbent Assay ◽  
Limit Of Detection ◽  
Deficiency Anemia ◽  
Enzyme Linked Immunosorbent Assay ◽  
Reactive Protein ◽  
Transient Rise ◽  
Juvenile Hemochromatosis ◽  
Serum Hepcidin ◽  
Clinical Conditions

Abstract We developed and validated the first serum enzyme-linked immunosorbent assay for hepcidin, the principal iron-regulatory hormone that has been very difficult to measure. In healthy volunteers, the 5% to 95% range of hepcidin concentrations was 29 to 254 ng/mL in men (n = 65) and 17 to 286 ng/mL in women (n = 49), with median concentrations 112 versus 65 (P < .001). The lower limit of detection was 5 ng/mL. Serum hepcidin concentrations in 24 healthy subjects correlated well with their urinary hepcidin (r = 0.82). Serum hepcidin appropriately correlated with serum ferritin (r = 0.63), reflecting the regulation of both proteins by iron stores. Healthy volunteers showed a diurnal increase of serum hepcidin at noon and 8 pm compared with 8 am, and a transient rise of serum hepcidin in response to iron ingestion. Expected alterations in hepcidin levels were observed in a variety of clinical conditions associated with iron disturbances. Serum hepcidin concentrations were undetectable or low in patients with iron deficiency anemia (ferritin < 10 ng/mL), iron-depleted HFE hemochromatosis, and juvenile hemochromatosis. Serum hepcidin concentrations were high in patients with inflammation (C-reactive protein > 10 mg/dL), multiple myeloma, or chronic kidney disease. The new serum hepcidin enzyme-linked immunosorbent assay yields accurate and reproducible measurements that appropriately reflect physiologic, pathologic, and genetic influences, and is informative about the etiology of iron disorders.

Immunoassay for Human Hepcidin in Blood

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 3839-3839
Author(s):  
Gordana Olbina ◽  
Ximena Pastorino ◽  
Domenico Girelli ◽  
Kimberly O. O’Brien ◽  
Elizabeta Nemeth ◽  
...  
Keyword(s):  
Healthy Volunteers ◽  
Limit Of Detection ◽  
Deficiency Anemia ◽  
Transient Rise ◽  
Juvenile Hemochromatosis ◽  
Heparin Plasma ◽  
Isotope Technique ◽  
Serum Hepcidin ◽  
Edta Plasma ◽  
Clinical Conditions

Abstract We developed and validated the first ELISA for hepcidin, the principal iron-regulatory hormone that has been very difficult to measure. The competitive ELISA uses a biotin-tagged hepcidin as a tracer to quantify hepcidin in serum, plasma or urine. In healthy volunteers, the 5–95% range of serum hepcidin concentrations was 29–254 ng/ml in men (n=65) and 17–286 ng/ml in women (n=49), with median concentrations 112 vs. 65, p<0.001. The lower limit of detection was 5 ng/ml. Serum, EDTA plasma and heparin plasma hepcidin measurements were in agreement (r> 0.98). Serum and urinary hepcidin concentrations in 24 healthy subjects correlated well (r=0.82). Serum hepcidin appropriately correlated with serum ferritin (r=0.63) reflecting the regulation of both proteins by iron stores. Healthy volunteers showed a diurnal increase of serum hepcidin at noon and 8pm compared to 8am, and a transient rise of serum hepcidin in response to iron ingestion. In 18 young healthy women, serum hepcidin correlated well (p<0.05) with intestinal iron absorption both from a food (r=0.49) and supplemental iron source (r=0.51), as determined by a stable iron isotope technique. In a variety of clinical conditions associated with iron disturbances we observed the appropriate changes in hepcidin concentrations: undetectable or low in patients with iron deficiency anemia (ferritin<10 ng/ml), treated HFE hemochromatosis, and juvenile hemochromatosis, and abnormally high in patients with inflammation (CRP>10 mg/dl), multiple myeloma, or chronic kidney disease. The new blood hepcidin ELISA yields accurate and reproducible measurements that appropriately reflect physiologic, pathologic and genetic influences, and is informative about the etiology of iron disorders.

Are Serum Hepcidin Levels Chronically Elevated in Collegiate Female Distance Runners?

2013 ◽  
Vol 23 (5) ◽  
pp. 513-521 ◽  
Author(s):  
Xiaoya Ma ◽  
Kaitlyn J. Patterson ◽  
Kayla M. Gieschen ◽  
Peter F. Bodary
Keyword(s):  
Iron Deficiency ◽  
Acute Exercise ◽  
Deficiency Anemia ◽  
Control Group ◽  
Distance Runners ◽  
Control Subjects ◽  
Transient Rise ◽  
Weekly Training ◽  
Serum Hepcidin ◽  
Clinical Conditions

The prevalence of iron deficiency tends to be higher in athletic populations, especially among endurancetrained females. Recent studies have provided evidence that the iron-regulating hormone hepcidin is transiently increased with acute exercise and suggest that this may contribute to iron deficiency anemia in athletes. The purpose of this study was to determine whether resting serum hepcidin is significantly elevated in highly trained female distance runners compared with a low exercise control group. Due to the importance of the monocyte in the process of iron recycling, monocyte expression of hepcidin was also measured. A single fasted blood sample was collected midseason from twenty female distance runners averaging 81.9 ± 14.2 km of running per week. Ten age-, gender-, and BMI-matched low-exercise control subjects provided samples during the same period using identical collection procedures. There was no difference between the runners (RUN) and control subjects (CON) for serum hepcidin levels (p = .159). In addition, monocyte hepcidin gene expression was not different between the two groups (p = .635). Furthermore, no relationship between weekly training volume and serum hepcidin concentration was evident among the trained runners. The results suggest that hepcidin is not chronically elevated with sustained training in competitive collegiate runners. This is an important finding because the current clinical conditions that link hepcidin to anemia include a sustained elevation in serum hepcidin. Nevertheless, additional studies are needed to determine the clinical relevance of the well-documented, transient rise in hepcidin that follows acute sessions of exercise.

scholarly journals POS0567 HEPCIDIN IS POTENTIAL BIOMARKER TO DISTINGUISH BETWEEN IRON DEFICIENCY ANEMIA AND ANEMIA OF INFLAMMATION IN RHEUMATOID ARTHRITIS

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 518.2-518
Author(s):  
E. Galushko ◽  
A. Semashko ◽  
A. Gordeev ◽  
A. Lila
Keyword(s):  
Rheumatoid Arthritis ◽  
Iron Deficiency ◽  
Iron Deficiency Anemia ◽  
Complete Blood Count ◽  
Deficiency Anemia ◽  
Reactive Protein ◽  
Potential Biomarker ◽  
Serum Hepcidin ◽  
Proper Diagnosis ◽  
Anemia Of Inflammation

Background:Anemia of inflammation (AI) and iron deficiency anemia (IDA) are the two most prevalent forms of anemia in patients with rheumatoid arthritis (RA). Diagnosis becomes challenging if AI is associated with true ID (AI/ID), as there is still a lack of a gold standard for differentiation between AI and AI/ID. However, as therapies to overcome anemia differ, proper diagnosis and understanding of underlying pathophysiological regulations are necessary.Objectives:The aim of the study was to evaluate the clinical efficiency of hepcidin, a key regulator of iron metabolism, in the diagnosis of IDA, as well as the differential diagnosis of AI/ID and AI in patients with RA.Methods:The study was undertaken 96 patients with RA, 67 of them were diagnosed anemia according to WHO criteria (104,3±21,4 g/l). Anemic patients and anemia-free patients with RA (n=29) were comparable (p>0.05) in age (44.4±14.8 and 49.8±9.3 years), disease duration (73.5±65.4 and 59.8±48.3 months) and DAS28 (6.3±1.6 and 5.9±1.9). All cases were subjected to following tests: complete blood count with peripheral smear, serum C-reactive protein, serum interleukin-6, iron studies, serum soluble transferrin receptor (sTfR), and serum hepcidin. Patients with RA and anemia were divided two groups: 25 patients with IDA and 42 - with AI. The AI cases were subdivided into pure AI and AI with coexistent ID (n=15).Results:The mean serum hepcidin concentration was significantly increased in pure AI patients (123.85±25.8 ng/mL) as compared to those in IDA patients (63.9±22.8 ng/mL, P < 0.05) and anemia-free patients with RA (88.1±39.09 ng/mL). Also, compared to pure AI patients [normal sTfR levels (<3 µg/mL)], the serum hepcidin concentration was reduced significantly in AI patients with ID [high sTfR levels (≥3 µg/mL)] with a mean of 79.0±23.97 ng/mL.Conclusion:Hepcidin measurement can provide a useful tool for differentiating AI from IDA and also help to identify an iron deficiency in AI patients. This might aid in the appropriate selection of therapy for these patients.Disclosure of Interests:None declared

scholarly journals Analysis of Specific Immunoglobulin G Subclass Antibodies for Serological Diagnosis of Echinococcosis by a Standard Enzyme-Linked Immunosorbent Assay

1998 ◽  
Vol 5 (5) ◽  
pp. 613-616 ◽  
Author(s):  
Felix Grimm ◽  
Friedrich E. Maly ◽  
Jian Lü ◽  
Roberto Llano
Keyword(s):  
Healthy Volunteers ◽  
Immunosorbent Assay ◽  
Immunoglobulin G ◽  
Serological Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Parasitic Infections ◽  
Igg Subclass ◽  
Specific Antibodies ◽  
Specific Igg4 ◽  
Antibody Levels

ABSTRACT The potential roles of specific antibodies of the different immunoglobulin G (IgG) subclasses in the serological diagnosis of cystic echinococcosis (CE) and alveolar echinococcosis (AE) were investigated by an enzyme-linked immunosorbent assay based on hydatid fluid as antigen. Specific antibodies of subclass 1 were found to be of major importance. In sera collected at the time of diagnosis (i.e., before any therapeutic intervention was initiated) they could be demonstrated in 14 of 15 sera from patients with CE and in all 12 sera from patients with AE. The most discriminatory and the most specific antibodies found in this study belonged to IgG subclass 4. Only one false-positive reaction was observed with 253 sera from healthy volunteers, and no cross-reactions occurred in 80 sera from patients with different parasitic infections. Specific IgG4 antibodies could be demonstrated in 61.0 to 66.7% (CE) or 47.6 to 66.7% (AE) of the cases. Antibody levels of IgG subclass 2 were elevated only moderately, and subclass 3 antibodies were detected in a few cases only. In addition, nonspecific reactions in sera of healthy volunteers or patients with other parasitic infections could partially be attributed to antibodies of subclasses 2 and 3.

Simulation model for predicting limit of detection and range of quantitation of competitive enzyme-linked immunosorbent assay

2007 ◽  
Vol 103 (5) ◽  
pp. 427-431 ◽  
Author(s):  
Dong Hwan Choi ◽  
Yoshio Katakura ◽  
Rieko Matsuda ◽  
Yuzuru Hayashi ◽  
Kazuaki Ninomiya ◽  
...  
Keyword(s):  
Simulation Model ◽  
Immunosorbent Assay ◽  
Limit Of Detection ◽  
Enzyme Linked Immunosorbent Assay

scholarly journals Aflatoxigenic and ochratoxigenic moulds and their toxins in tree nuts on Serbian market

2020 ◽  
pp. 9-18
Author(s):  
Irena Rakic ◽  
Gordana Dimic ◽  
Marija Skrinjar ◽  
Suncica Kocic-Tanackov
Keyword(s):  
Aspergillus Niger ◽  
Ochratoxin A ◽  
Immunosorbent Assay ◽  
Alternaria Alternata ◽  
Rhizopus Oryzae ◽  
Limit Of Detection ◽  
Average Concentration ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mean Values ◽  
Tree Nuts

In this study, moulds and mycotoxins presence in different tree nuts were investigated. The results showed that all of the 25 samples were contaminated with moulds. Mean values of total mould count varied from 1-4.9 cfu per grain. The most frequent species in hazelnut samples were Rhizopus oryzae (32.2%) and Aspergillus niger (28.9%). In walnuts A. niger (75.6%), in cashews also A. niger (42.4%) while in pistachio samples Alternaria alternata (20.7%), and Cladosporium cladosporioides (20.7%) were the most dominant. Rhizopus oligosporus was the only identified species in all almond samples (100%). Using Enzyme Linked Immunosorbent Assay (ELISA), the presence of total aflatoxins and ochratoxin A was examinated. In all analyzed samples, levels of ochratoxin A were below the limit of detection. Total aflatoxins were detected only in walnut samples with average concentration of 7.1 ?g/kg.

A Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay for Determination of Homoharringtonine

Planta Medica ◽  
2018 ◽  
Vol 84 (14) ◽  
pp. 1038-1044 ◽  
Author(s):  
Benyakan Pongkitwitoon ◽  
Seiichi Sakamoto ◽  
Rika Nagamitsu ◽  
Waraporn Putalun ◽  
Hiroyuki Tanaka ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Myeloid Leukemia ◽  
High Performance ◽  
Sodium Periodate ◽  
Limit Of Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Study Support ◽  
Mediated Oxidation

AbstractHomoharringtonine (HHT), also known as omacetaxine, is a natural compound found in the genus Cephalotaxus and is a promising pharmaceutical drug used for the treatment of chronic or accelerated phase chronic myeloid leukemia. As a tool for the quantitative determination of HHT, a specific monoclonal antibody against HHT (MAb 6A1) was generated by conjugates prepared via sodium periodate-mediated oxidation. The developed indirect competitive enzyme-linked immunosorbent assay (icELISA) using MAb 6A1 was found to be highly specific and sensitive with a limit of detection for HHT of 48.8 ng/mL. Validation assays to evaluate precision and accuracy of the method were conducted by the use of intra- and inter-assay analysis, recovery test, and comparison analysis between the amounts of HHT determined by ELISA and high-performance liquid chromatography. These results revealed that the established icELISA using MAb 6A1 is specific, sensitive, and reliable enough to be applied to the qualitative analysis for HHT. Furthermore, the results of this study support the usefulness of sodium periodate as a reagent for the conjugation between Cephalotaxus alkaloids and proteins for producing specific antibodies.

Measurement of serum amyloid A1 (SAA1), a major isotype of acute phase SAA

2006 ◽  
Vol 44 (1) ◽  
Author(s):  
Yuanyuan Xu ◽  
Toshiyuki Yamada ◽  
Takahiko Satoh ◽  
Yasuaki Okuda
Keyword(s):  
Acute Phase ◽  
Healthy Subjects ◽  
Acute Phase Proteins ◽  
Serum Amyloid ◽  
Enzyme Linked Immunosorbent Assay ◽  
Reactive Protein ◽  
Amyloid A ◽  
Amyloid Deposits ◽  
Clinical Conditions ◽  
Kinetics Of

AbstractSerum amyloid A (SAA), a plasma precursor of reactive amyloid deposits, is a multigene product. SAA1 and SAA2, with primary structures that are 93% identical (98 of 104 amino acids), behave as acute phase proteins, as demonstrated by their increasing levels in plasma. Heretofore, it has been understood that SAA1 predominates and functions as an isotype in plasma. However, accurate measurements differentiating the two isotypes have not been reported. In this study, using monoclonal antibodies specific for SAA1, we developed an enzyme-linked immunosorbent assay (ELISA) for SAA1. The levels and ratios of SAA1 in total SAA (TSAA) were investigated in healthy subjects and patients with rheumatoid arthritis (RA). The SAA1/TSAA ratio was 74±12% and 77±12% in healthy subjects and RA patients, respectively. In RA patients, the ratios were not influenced by SAA1 genotype, which has been proposed to affect plasma SAA values. The kinetics of SAA1 in inflamed patients undergoing hemodialysis was found to be parallel with total SAA and C-reactive protein. Finally, this study confirmed that SAA1 is a major isotype of acute phase SAA and may determine total SAA values. This specific assay could be used in the evaluation of SAA behavior in several clinical conditions.

scholarly journals Development of an antigen-capture enzyme-linked immunosorbent assay for Clostridium perfringens beta2-toxin in porcine feces and the neonatal piglet intestine

2012 ◽  
Vol 24 (5) ◽  
pp. 895-902 ◽  
Author(s):  
Jasmina Kircanski ◽  
Douglas Hodgins ◽  
Glenn Soltes ◽  
Yanlong Pei ◽  
Valeria R. Parreira ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Protease Activity ◽  
Limit Of Detection ◽  
Intestinal Content ◽  
Enzyme Linked Immunosorbent Assay ◽  
Neonatal Piglet ◽  
Antigen Capture ◽  
Field Samples ◽  
Intestinal Contents ◽  
Beta2 Toxin

An enzyme-linked immunosorbent assay (ELISA) was developed for detection and quantitation of beta2-toxin in neonatal piglet intestinal contents. Polystyrene plates were coated with polyclonal capture antibodies prepared against consensus recombinant beta2-toxin. The ELISA was developed using consensus recombinant beta2-toxin, atypical recombinant beta2-toxin, purified consensus native beta2-toxin, and field samples of neonatal porcine intestinal contents. Captured antigen was detected using a horseradish peroxidase–labeled monoclonal antibody against consensus recombinant beta2-toxin. The limit of detection of the ELISA for consensus beta2-toxin was between 2.0 and 3.5 ng/ml. The ELISA detected atypical recombinant beta2-toxin only weakly. Optical density was protein concentration dependent. The test confirmed differences between consensus and atypical recombinant beta2-toxin, but similar results obtained when testing pure consensus recombinant beta2-toxin and native beta2-toxin. Results obtained from intestinal content samples, particularly from the small intestine, were highly inconsistent and suggested variable protease activity. Addition of protease inhibitors partially prevented degradation of the toxin; however, sample processing at low temperature, at a lower pH (citrate buffer with 5% of bovine serum albumin, pH 6.1), and “cold incubation” of applied antigens abolished protease activity. The recombinant toxin was preserved in spiked intestinal samples by freezing at −70°C, suggesting that necropsy samples can be stored frozen for periodic testing. With appropriate sample preparation, antigen-capture ELISA can detect beta2-toxin in the intestinal content and feces of neonatal piglets.

scholarly journals High Sensitivity C-Reactive Protein sebagai Parameter Diagnostik dan Prediktor Luaran Sepsis pada Anak yang Menderita Systemic Inflammatory Response Syndrome

Sari Pediatri ◽  
2016 ◽  
Vol 16 (4) ◽  
pp. 278
Author(s):  
Sofni Sarmen ◽  
Mayetti Mayetti ◽  
Hafni Bachtiar
Keyword(s):  
Systemic Inflammatory Response Syndrome ◽  
Inflammatory Response ◽  
Immunosorbent Assay ◽  
High Sensitivity ◽  
Systemic Inflammatory Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
C Reactive Protein ◽  
Reactive Protein ◽  
Hs Crp ◽  
Elisa Data

Latar belakang. Sepsis merupakan salah satu penyebab morbiditas dan mortalitas pada anak. Diagnosissepsis ditegakkan berdasarkan gejala Systemic Inflammatory Response Syndrome (SIRS) dan penemuan bakteripada kultur darah. Kultur bakteri darah memiliki sensitifitas yang rendah dan membutuhkan waktu yanglama sehingga sering menyebabkan terjadinya overdiagnosis dan overtreatment. C-reactive protein adalahreaktan fase akut yang kadarnya meningkat pada keadaan infeksi. High sensitivity C-reactive protein (hs-CRP) adalah metode yang lebih sensitif untuk mengukur kadar CRP dalam jumlah kecil.Tujuan. Mengetahui peran hs-CRP sebagai parameter diagnostik dan prediktor luaran sepsis pada anakyang menderita SIRS.Metode. Penelitian uji diagnostik dengan desain potong lintang terhadap 85 anak dengan gejala SIRS berusia1 bulan sampai dengan 15 tahun dan dirawat di bangsal anak RS.Dr.M.Djamil Padang sejak Juni sampaiNovember 2012. Pemeriksaan hs-CRP dilakukan dengan metode enzyme-linked immunosorbent assay (ELISA).Data dianalisis dengan SPSS serta dilakukan uji diagnostik. Baku emas sepsis adalah biakan darah.Hasil. Cut off point hs-CRP untuk menentukan sepsis adalah 15,55 ng/ml, (sensitivitas 90,9% dan spesivisitas53,8%). Kadar rata-rata hs-CRP meningkat sesuai dengan beratnya penyakit.Kesimpulan. High sensitivity C-reactive protein dapat dijadikan sebagai parameter diagnostik sepsis padapasien SIRS dengan cut off point 15,55 ng/ml, serta dapat dipakai sebagai prediktor luaran sepsis.

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