Optimization of a procedure to accurately detect equine TNFα in serum samples.
Abstract: The systemic component of chronic inflammatory diseases may lead to clinical complications. High levels of TNFα, a pro-inflammatory cytokine, are found in human patients with COPD and asthma. Horses are also susceptible to an array of chronic inflammatory disorders possibly associated with systemic inflammation, including respiratory diseases. Currently, there is no commercially available ELISA validated to assess TNFα in equine serum samples. Moreover, the reported normal mean concentration of serum TNFα in horses vary greatly. Hence, we sought to optimize and validate a procedure to quantify this cytokine in equine serum samples using a sandwich ELISA. Our results indicate that the nature of diluent buffers greatly impact the detection of TNFα in equine serum samples as its quantification increased in some cases from non-detectable levels to the ng/ml range. Linearity assays performed with serum samples from six animals serially diluted in four different buffers showed that serum matrix interference was animal-dependant. The specificity of TNFα detection was also assessed. Our optimized assay conditions were validated by quantifying levels of TNFα in serum samples from normal horses and horses affected with chronic pulmonary disease (heaves).
Copyright © 2010 Elsevier B.V. All rights reserved.
Publication Date: 2010-07-06 PubMed ID: 20674987DOI: 10.1016/j.vetimm.2010.06.018Google Scholar: Lookup
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- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
- Validation Study
- Asthma
- Chronic Obstructive Pulmonary Disease
- Clinical Study
- Cytokines
- Diagnosis
- Diagnostic Technique
- Disease Diagnosis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Equine Diseases
- Equine Health
- Horses
- Immunology
- In Vivo
- Inflammation
- Laboratory Methods
- Physiology
- Respiratory Disease
- Serum
- Tumor Necrosis Factor
- Veterinary Medicine
- Veterinary Research
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The study aims to improve and authenticate a method to measure the levels of TNFα, a bodily chemical linked to inflammation, in the blood of horses. Evidence shows that a successful assay was developed enabling enhanced detection of TNFα in horse serum.
Objective of Study
- The main goal of the study was to develop an efficient and reliable method to measure the levels of TNFα, an inflammatory marker, in horse blood serum. This optimization is critical for understanding and treating chronic inflammatory conditions in horses, which in many cases mimic human conditions like asthma and COPD.
Methods and Process
- The researchers used a sandwich ELISA (enzyme-linked immunosorbent assay) – a common laboratory technique for detecting substances in liquid samples. However, no such commercially available ELISA is validated for assessing TNFα in equine samples.
- It was observed that different diluent buffers could greatly influence the detection of TNFα in horse serum. In fact, depending on the buffer, the levels of TNFα went from undetectable levels to the ng/ml range.
- The linearity, which describes how much the assay response increases with an increase in the analyte, was checked with samples from six different animals. This was done by diluting the samples serially in four different buffers. The results showed that the serum’s matrix interference was animal-dependant.
Findings and Validation
- The researchers also checked the specificity of TNFα detection to ensure that the assay was recognizing and measuring the cytokine accurately.
- The newfound optimized ELISA conditions were validated using serum samples collected from both healthy horses and those with chronic pulmonary disease, known as heaves.
Implications of the Study
- This validated assay procedure can significantly aid in understanding chronic inflammatory disorders in horses and potentially building therapeutic interventions. The outcome could also offer valuable insights for similar conditions in humans.
Cite This Article
APA
Lavoie-Lamoureux A, Maghni K, Lavoie JP.
(2010).
Optimization of a procedure to accurately detect equine TNFα in serum samples.
Vet Immunol Immunopathol, 138(1-2), 118-123.
https://doi.org/10.1016/j.vetimm.2010.06.018 Publication
Researcher Affiliations
- Faculté de Médecine Vétérinaire, Université de Montréal, 3200 rue Sicotte, St-Hyacinthe, J2S2M2 Québec, Canada. Anouk.lavoie.lamoureux@umontreal.ca
MeSH Terms
- Animals
- Blood Chemical Analysis / methods
- Blood Chemical Analysis / veterinary
- Buffers
- Enzyme-Linked Immunosorbent Assay / methods
- Enzyme-Linked Immunosorbent Assay / veterinary
- Horse Diseases / blood
- Horse Diseases / immunology
- Horses / blood
- Horses / immunology
- Humans
- Inflammation Mediators / blood
- Lung Diseases / blood
- Lung Diseases / immunology
- Lung Diseases / veterinary
- Reproducibility of Results
- Tumor Necrosis Factor-alpha / blood
Citations
This article has been cited 5 times.- Mukhopadhyay A, Cook SR, SanMiguel P, Ekenstedt KJ, Taylor SD. TLR4 and MD2 variation among horses with differential TNFα baseline concentrations and response to intravenous lipopolysaccharide infusion. Sci Rep 2023 Jan 27;13(1):1486.
- Anderson MJ, Ibrahim AS, Cooper BR, Woolcock AD, Moore GE, Taylor SD. Effects of administration of ascorbic acid and low-dose hydrocortisone after infusion of sublethal doses of lipopolysaccharide to horses. J Vet Intern Med 2020 Nov;34(6):2710-2718.
- Bamford NJ, Potter SJ, Baskerville CL, Harris PA, Bailey SR. Influence of dietary restriction and low-intensity exercise on weight loss and insulin sensitivity in obese equids. J Vet Intern Med 2019 Jan;33(1):280-286.
- Schnabel CL, Steinig P, Koy M, Schuberth HJ, Juhls C, Oswald D, Wittig B, Willenbrock S, Murua Escobar H, Pfarrer C, Wagner B, Jaehnig P, Moritz A, Feige K, Cavalleri JM. Immune response of healthy horses to DNA constructs formulated with a cationic lipid transfection reagent. BMC Vet Res 2015 Jun 23;11:140.
- Richard EA, Depecker M, Defontis M, Leleu C, Fortier G, Pitel PH, Couroucé-Malblanc A. Cytokine concentrations in bronchoalveolar lavage fluid from horses with neutrophilic inflammatory airway disease. J Vet Intern Med 2014 Nov-Dec;28(6):1838-44.
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