Validity and application of immunoturbidimetric and enzyme-linked immunosorbent assays for the measurement of adiponectin concentration in ponies.
Abstract: Circulating adiponectin concentrations were lower in ponies with a history of endocrinopathic laminitis and in nonlaminitic ponies that subsequently developed laminitis. The assays used in these studies have been discontinued or are no longer valid. Objective: (1) to determine the validity of immunoturbidimetric (IT) and enzyme linkedimmunosorbent (ELISA) assays for equine total and high molecular weight (HMW) [adiponectin] measurement and (2) to investigate the association between [adiponectin] measured using these assays and endocrinopathic laminitis. Methods: Method validation and cohort study. Methods: Accuracy and precision of IT and ELISA assays for measuring total (TAC) and HMW (HMWAC) [adiponectin] were determined. Using the IT assay, the effects of anti-coagulant and storage temperature were assessed, TAC was measured in previously laminitic (PL) and never laminitic (NL) ponies (n = 6/group). Comparison with a previously validated radioimmunoassay was made in NL ponies (n = 223). Association between TAC and subsequent laminitis development in NL ponies was investigated using univariable logistic regression and ROC curve analysis. Results: The IT assay was precise and demonstrated good agreement with the previously validated radioimmunoassay. TAC was significantly (P<0.01) lower in PL (mean ± s.d. 8.9 ± 2.9 μg/mL) compared to NL (24.2 ± 11.8 μg/mL) ponies and in NL ponies that developed laminitis within 12 months (median 4.8 μg/mL; IQR 2.65-13.4 μg/mL) compared to those that remained nonlaminitic (19.9 μg/mL; 9.95-31.5 μg/mL). TAC was significantly (P = 0.01) associated with laminitis occurrence within 12 months. Use of the area under the ROC curve to distinguish animals that did and did not develop laminitis showed good accuracy (0.76). None of the ELISA methods validated satisfactorily. Conclusions: Laminitis risk is based on data from ponies in one region. Conclusions: The IT method is suitable for measurement of equine TAC. TAC is lower in ponies with previous or future laminitis. The ELISA methods are not suitable for measurement of equine HMWAC or TAC.
© 2018 EVJ Ltd.
Publication Date: 2018-05-17 PubMed ID: 29679396DOI: 10.1111/evj.12960Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Validation Study
- Clinical Pathology
- Clinical Study
- Diagnosis
- Diagnostic Technique
- Disease Diagnosis
- Disease Etiology
- Disease Treatment
- Endocrine System
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Epidemiology
- Equine Health
- Horses
- Immunoassay
- Immunology
- In Vivo
- Laboratory Methods
- Lameness
- Metabolic Health
- Pony
- 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.
This research investigates the validity of two methods, Immunoturbidimetric and enzyme-linked immunosorbent assays, for measuring the concentration of adiponectin – a protein hormone – in ponies. The study also explores the connection between adiponectin and endocrinopathic laminitis, a hoof disease in horses.
Methodology
- The researchers performed tests to ascertain the accuracy and precision of both the immunoturbidimetric (IT) and enzyme-linked immunosorbent (ELISA) assays for measuring total and high molecular weight adiponectin.
- The team conducted experiments to assess the effects of the presence of an anticoagulant and storage temperature on the IT assay.
- They measured the Total Adiponectin Concentration (TAC) in two groups of ponies; those that previously had endocrinopathic laminitis (PL) and those that never had the hoof disorder (NL).
- The performance of the assays were compared against an already validated method, the radioimmunoassay.
- Lastly, they made strides to identify the association between TAC and the development of endocrinopathic laminitis in NL ponies.
Results
- The IT was accurate and aligned well with the previously validated radioimmunoassay. However, the ELISA methods were not properly validated.
- TAC measured was significantly lower in ponies that had suffered laminitis and in those NL ponies that later showed symptoms within 12 months.
- Adequate accuracy was found when the area under the Receiver Operating Characteristic (ROC) curve was used to differentiate the animals that did and those that did not contract laminitis.
Conclusions
- The data indicated laminitis risk is based on ponies from one region.
- The Immunoturbidimetric method was valid for measuring equine TAC. However, the ELISA methods were not suited, neither for TTAC nor for high molecular weight adiponectin.
- Ponies with lower TAC either had a history of laminitis or subsequently developed the ailment.
These findings highlight the relevance of adiponectin as a potential biomarker for predicting the risk of endocrinopathic laminitis in equines. Further research should establish benchmarks for normal adiponectin levels to better manage horses at risk for developing this disorder.
Cite This Article
APA
Menzies-Gow NJ, Knowles EJ, Rogers I, Rendle DI.
(2018).
Validity and application of immunoturbidimetric and enzyme-linked immunosorbent assays for the measurement of adiponectin concentration in ponies.
Equine Vet J, 51(1), 33-37.
https://doi.org/10.1111/evj.12960 Publication
Researcher Affiliations
- Department of Clinical Sciences and Services, Royal Veterinary College, North Mymms, Hertfordshire, UK.
- Department of Clinical Sciences and Services, Royal Veterinary College, North Mymms, Hertfordshire, UK.
- Rainbow Equine Lab, Malton, North Yorkshire, UK.
- Rainbow Equine Lab, Malton, North Yorkshire, UK.
MeSH Terms
- Adiponectin / blood
- Adiponectin / chemistry
- Animals
- Anticoagulants / therapeutic use
- Biomarkers / blood
- Cohort Studies
- Enzyme-Linked Immunosorbent Assay / standards
- Enzyme-Linked Immunosorbent Assay / veterinary
- Female
- Foot Diseases / blood
- Foot Diseases / veterinary
- Hoof and Claw
- Horse Diseases / blood
- Horses / blood
- Immunoturbidimetry / standards
- Immunoturbidimetry / veterinary
- Logistic Models
- Molecular Weight
- ROC Curve
- Radioimmunoassay / standards
- Radioimmunoassay / veterinary
- Reproducibility of Results
- Risk Factors
- Temperature
- Time Factors
Grant Funding
- Mellon Trust
- MARS PetCare UK
- Rainbow Equine Lab
Citations
This article has been cited 4 times.- Karikoski NP, Box JR, Mykkänen AK, Kotiranta VV, Raekallio MR. Variation in insulin response to oral sugar test in a cohort of horses throughout the year and evaluation of risk factors for insulin dysregulation.. Equine Vet J 2022 Sep;54(5):905-913.
- Wise JC, Wilkes EJA, Raidal SL, Xie G, Crosby DE, Hale JN, Hughes KJ. Interobserver and intraobserver reliability for 2 grading systems for gastric ulcer syndrome in horses.. J Vet Intern Med 2021 Jan;35(1):571-579.
- Fitzgerald DM, Anderson ST, Sillence MN, de Laat MA. The cresty neck score is an independent predictor of insulin dysregulation in ponies.. PLoS One 2019;14(7):e0220203.
- Durham AE, Frank N, McGowan CM, Menzies-Gow NJ, Roelfsema E, Vervuert I, Feige K, Fey K. ECEIM consensus statement on equine metabolic syndrome.. J Vet Intern Med 2019 Mar;33(2):335-349.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
