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Home / Equine Research Bank / J Vet Intern Med / Evaluation of a Chemiluminescent Immunoassay for Measurement...
Journal of veterinary internal medicine2017; 31(2); 568-574; doi: 10.1111/jvim.14657

Evaluation of a Chemiluminescent Immunoassay for Measurement of Equine Insulin.

Abstract: Many diagnostic tests for insulin dysregulation use reference intervals established with an insulin radioimmunoassay (RIA) that is no longer available. A chemiluminescent immunoassay (CLIA) is commonly used for the measurement of serum insulin concentration in clinical practice but requires further validation, especially at clinically relevant reference intervals. Objective: To evaluate the CLIA for measurement of equine insulin and compare it to the previously validated, but now unavailable RIA. Methods: Equine serum samples (n = 78) from clinical and experimental studies. Methods: In this experimental study, performance of the CLIA was evaluated using standard variables, including comparison with the RIA. Continuous and binary outcomes were analyzed. Results: The CLIA showed good intra-assay (coefficient of variation [CV], 1.8-2.4%) and interassay (CV, 3-7.1%) precision. Acceptable recovery on dilution (100 ± 10%) was achieved only at dilutions <1:1. Recovery on addition was acceptable. Comparison of the CLIA and RIA showed strong positive correlation (r = 0.91-0.98), with fixed and proportional bias. At 3 diagnostic cutoffs, sensitivity of CLIA compared with RIA ranged from 67 to 100% and specificity from 96 to 100%. Conclusions: The CLIA is a highly repeatable assay which is suitable for within- and between-horse comparisons. Dilution of high concentration samples should be performed with charcoal-stripped serum (CSS) and at the lowest dilution factor possible. At concentrations commonly used for diagnosis of insulin dysregulation (≤100 μIU/mL), results from the CLIA tend to be lower than from the RIA and should be interpreted accordingly. Further standardization of equine insulin assays is required.
Copyright © 2017 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.
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Publication Date: 2017-01-26 PubMed ID: 28124389PubMed Central: PMC5354037DOI: 10.1111/jvim.14657Google Scholar: Lookup
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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 article focuses on evaluating the effectiveness of a Chemiluminescent Immunoassay (CLIA) in measuring equine insulin levels as it is compared to a previously validated but now unavailable technique known as Insulin Radioimmunoassay (RIA). The study finds that while the CLIA yields reliable results, further standardization of equine insulin assays is required.

Methodology

  • Researchers conducted an experimental study using equine serum samples obtained from previous clinical and experimental trials. The total number of samples analyzed remained 78.
  • They evaluated the performance of the CLIA in measuring equine insulin using several standard variables and then compared these results to those generated by RIA.

Results

  • The Chemiluminescent Immunoassay technique exhibited strong intra-assay and interassay accuracy, with precision defined in terms of coefficient variation (CV). These precision levels ranged between 1.8 and 2.4% for intra-assay and between 3 and 7.1% for interassay, indicating a high level of repeatability.
  • This technique also achieved an acceptable recovery rate on dilution but only at dilutions below 1:1. The recovery on direct addition was also deemed to be acceptable.
  • Strong positive correlation was observed between the results of the CLIA and RIA (r = 0.91-0.98). However, this comparison also revealed fixed and proportional biases.
  • The sensitivity of the CLIA compared to the RIA fluctuated between 67 to 100%, and specificity ranged from 96 to 100% across three diagnostic cut-offs.

Conclusions

  • From the results obtained, the study concluded that the CLIA method, due to its high repeatability, suitability for within-horse and between-horse comparisons and reasonable sensitivity, proved to be a viable technique for measuring equine insulin at clinically relevant levels.
  • However, diluting samples with high concentrations should be achieved using a charcoal-stripped serum (CSS) and at the lowest possible dilution factor.
  • It should be noted that at lower insulin concentrations, the result from the CLIA is lower than from the RIA, indicating some degree of underestimation. This suggests the need for adjusting the interpretation of results accordingly.
  • The researchers ultimately recommended further standardization of equine insulin assays in order to attain more accurate and consistent results in clinical diagnosis.

Cite This Article

APA
Carslake HB, Pinchbeck GL, McGowan CM. (2017). Evaluation of a Chemiluminescent Immunoassay for Measurement of Equine Insulin. J Vet Intern Med, 31(2), 568-574. https://doi.org/10.1111/jvim.14657

Publication

Journal of veterinary internal medicine
ISSN: 1939-1676
NlmUniqueID: 8708660
Country: United States
Language: English
Volume: 31
Issue: 2
Pages: 568-574

Researcher Affiliations

Carslake, H B
  • Institute of Aging and Chronic Disease, Faculty of Health and Life Sciences, University of Liverpool, Neston, Wirral, UK.
Pinchbeck, G L
  • Institute of Infection and Global Health, Faculty of Health and Life Sciences, University of Liverpool, Neston, Wirral, UK.
McGowan, C M
  • Institute of Aging and Chronic Disease, Faculty of Health and Life Sciences, University of Liverpool, Neston, Wirral, UK.

MeSH Terms

  • Animals
  • Horses / blood
  • Immunoassay / veterinary
  • Insulin / blood
  • Luminescent Measurements / methods
  • Luminescent Measurements / veterinary
  • Radioimmunoassay / methods
  • Radioimmunoassay / veterinary
  • Reproducibility of Results
  • Sensitivity and Specificity

References

This article includes 18 references
  1. Asplin KE, Sillence MN, Pollitt CC, McGowan CM. Induction of laminitis by prolonged hyperinsulinaemia in clinically normal ponies.. Vet J 2007 Nov;174(3):530-5.
    pubmed: 17719811doi: 10.1016/j.tvjl.2007.07.003google scholar: lookup
  2. Carter RA, Treiber KH, Geor RJ, Douglass L, Harris PA. Prediction of incipient pasture-associated laminitis from hyperinsulinaemia, hyperleptinaemia and generalised and localised obesity in a cohort of ponies.. Equine Vet J 2009 Feb;41(2):171-8.
    pubmed: 19418747doi: 10.2746/042516408x342975google scholar: lookup
  3. Jeffcott LB, Field JR, McLean JG, O'Dea K. Glucose tolerance and insulin sensitivity in ponies and Standardbred horses.. Equine Vet J 1986 Mar;18(2):97-101.
    pubmed: 3516677doi: 10.1111/j.2042-3306.1986.tb03556.xgoogle scholar: lookup
  4. Karikoski NP, McGowan CM, Singer ER, Asplin KE, Tulamo RM, Patterson-Kane JC. Pathology of Natural Cases of Equine Endocrinopathic Laminitis Associated With Hyperinsulinemia.. Vet Pathol 2015 Sep;52(5):945-56.
    pubmed: 25232034doi: 10.1177/0300985814549212google scholar: lookup
  5. Treiber KH, Kronfeld DS, Hess TM, Byrd BM, Splan RK, Staniar WB. Evaluation of genetic and metabolic predispositions and nutritional risk factors for pasture-associated laminitis in ponies.. J Am Vet Med Assoc 2006 May 15;228(10):1538-45.
    pubmed: 16677122doi: 10.2460/javma.228.10.1538google scholar: lookup
  6. nMercodian. Mercodia Equine insulin ELISA. Directions for Use Version 4.0. Available at: https://www.mercodia.se/assets/upload/files/DfU/Equine/Insulin%20Equine%20version%204_0.pdf 2016. Accessed July 23, 2016.
  7. Eiler H, Frank N, Andrews FM, Oliver JW, Fecteau KA. Physiologic assessment of blood glucose homeostasis via combined intravenous glucose and insulin testing in horses.. Am J Vet Res 2005 Sep;66(9):1598-604.
    pubmed: 16261835doi: 10.2460/ajvr.2005.66.1598google scholar: lookup
  8. Tinworth KD, Wynn PC, Boston RC, Harris PA, Sillence MN, Thevis M, Thomas A, Noble GK. Evaluation of commercially available assays for the measurement of equine insulin.. Domest Anim Endocrinol 2011 Aug;41(2):81-90.
    pubmed: 21741576doi: 10.1016/j.domaniend.2011.05.001google scholar: lookup
  9. Banse HE, McCann J, Yang F, Wagg C, McFarlane D. Comparison of two methods for measurement of equine insulin.. J Vet Diagn Invest 2014 Jul;26(4):527-530.
    pubmed: 24928598doi: 10.1177/1040638714536560google scholar: lookup
  10. Morgan RA, Keen JA, McGowan CM. Treatment of equine metabolic syndrome: A clinical case series.. Equine Vet J 2016 Jul;48(4):422-6.
    pubmed: 25808563doi: 10.1111/evj.12445google scholar: lookup
  11. Herbert V, Lau KS, Gottlieb CW, Bleicher SJ. Coated charcoal immunoassay of insulin.. J Clin Endocrinol Metab 1965 Oct;25(10):1375-84.
    pubmed: 5320561doi: 10.1210/jcem-25-10-1375google scholar: lookup
  12. Deming WE. Statistical Adjustment of Data. New York: John Wiley and Sons; 1943.
  13. Bland JM, Altman DG. Statistical methods for assessing agreement between two methods of clinical measurement.. Lancet 1986 Feb 8;1(8476):307-10.
    pubmed: 2868172
  14. Borer-Weir KE, Bailey SR, Menzies-Gow NJ, Harris PA, Elliott J. Evaluation of a commercially available radioimmunoassay and species-specific ELISAs for measurement of high concentrations of insulin in equine serum.. Am J Vet Res 2012 Oct;73(10):1596-602.
    pubmed: 23013186doi: 10.2460/ajvr.73.10.1596google scholar: lookup
  15. Öberg J, Bröjer J, Wattle O. Evaluation of an equine‐optimized enzyme‐linked immunosorbent assay for serum insulin measurement and stability study of equine serum insulin. Comp Clin Path 2011;21:1291–1300.
  16. Tinworth KD, Wynn PC, Harris PA. Optimising the Siemens Coat‐A‐Count radioimmunoassay to measure insulin in equine plasma. J Equine Vet Sci 2009;29:411–413.
  17. Ho EN, Wan TS, Wong AS, Lam KK, Stewart BD. Doping control analysis of insulin and its analogues in equine plasma by liquid chromatography-tandem mass spectrometry.. J Chromatogr A 2008 Aug 8;1201(2):183-90.
    pubmed: 18501368doi: 10.1016/j.chroma.2008.04.060google scholar: lookup
  18. Rodríguez-Cabaleiro D, Van Uytfanghe K, Stove V, Fiers T, Thienpont LM. Pilot study for the standardization of insulin immunoassays with isotope dilution liquid chromatography/tandem mass spectrometry.. Clin Chem 2007 Aug;53(8):1462-9.
    pubmed: 17586588doi: 10.1373/clinchem.2007.088393google scholar: lookup

Citations

This article has been cited 15 times.
  1. Delarocque J, Feige K, Carslake HB, Durham AE, Fey K, Warnken T. Development of a Web App to Convert Blood Insulin Concentrations among Various Immunoassays Used in Horses.. Animals (Basel) 2023 Aug 24;13(17).
    doi: 10.3390/ani13172704pubmed: 37684968google scholar: lookup
  2. Gupta A, Bhat HR, Singh UP. Discovery of imeglimin-inspired novel 1,3,5-triazine derivatives as antidiabetic agents in streptozotocin-induced diabetes in Wistar rats via inhibition of DPP-4.. RSC Med Chem 2023 Aug 16;14(8):1512-1536.
    doi: 10.1039/d3md00085kpubmed: 37593577google scholar: lookup
  3. Clark BL, Bamford NJ, Stewart AJ, McCue ME, Rendahl A, Bailey SR, Bertin FR, Norton EM. Evaluation of an HMGA2 variant contribution to height and basal insulin concentrations in ponies.. J Vet Intern Med 2023 May-Jun;37(3):1186-1192.
    doi: 10.1111/jvim.16723pubmed: 37148171google scholar: lookup
  4. Go YY, Hazard NW, Balasuriya UBR, Chapman AM, Fitton NS, Kenéz Á, Andrews FM. Clinical evaluation of the Immulite® 1000 chemiluminescent immunoassay for measurement of equine serum insulin.. Front Vet Sci 2023;10:1018230.
    doi: 10.3389/fvets.2023.1018230pubmed: 37051514google scholar: lookup
  5. Berryhill EH, Urbina NS, Marton S, Vernau W, Alonso FH. Validation and method comparison for a point-of-care lateral flow assay measuring equine whole blood insulin concentrations.. J Vet Diagn Invest 2023 Mar;35(2):124-131.
    doi: 10.1177/10406387221142288pubmed: 36482705google scholar: lookup
  6. Williams NJ, Furr M, Navas de Solis C, Campolo A, Davis M, Lacombe VA. Investigating the Relationship Between Cardiac Function and Insulin Sensitivity in Horses: A Pilot Study.. Front Vet Sci 2022;9:899951.
    doi: 10.3389/fvets.2022.899951pubmed: 35873691google scholar: lookup
  7. Kinsella HM, Hostnik LD, Snyder HA, Mazur SE, Kamr AM, Burns TA, Mossbarger JC, Toribio RE. Comparison of insulin sensitivity between healthy neonatal foals and horses using minimal model analysis.. PLoS One 2022;17(1):e0262584.
    doi: 10.1371/journal.pone.0262584pubmed: 35030228google scholar: lookup
  8. Stokes SM, Stefanovski D, Bertin FR, Medina-Torres CE, Belknap JK, van Eps AW. Plasma amino acid concentrations during experimental hyperinsulinemia in 2 laminitis models.. J Vet Intern Med 2021 May;35(3):1589-1596.
    doi: 10.1111/jvim.16095pubmed: 33704816google scholar: lookup
  9. Carslake HB, Pinchbeck GL, McGowan CM. Equine metabolic syndrome in UK native ponies and cobs is highly prevalent with modifiable risk factors.. Equine Vet J 2021 Sep;53(5):923-934.
    doi: 10.1111/evj.13378pubmed: 33128277google scholar: lookup
  10. d' Fonseca NMM, Gibson CME, van Doorn DA, de Ruijter-Villani M, Stout TAE, Roelfsema E. Effect of long-term overfeeding of a high-energy diet on glucose tolerance in Shetland pony mares.. J Vet Intern Med 2020 May;34(3):1339-1349.
    doi: 10.1111/jvim.15788pubmed: 32374454google scholar: lookup
  11. Kritchevsky JE, Muir GS, Leschke DHZ, Hodgson JK, Hess EK, Bertin FR. Blood glucose and insulin concentrations after alpha-2-agonists administration in horses with and without insulin dysregulation.. J Vet Intern Med 2020 Mar;34(2):902-908.
    doi: 10.1111/jvim.15747pubmed: 32100334google scholar: lookup
  12. Leschke DH, Muir GS, Hodgson JK, Coyle M, Horn R, Bertin FR. Immunoreactive insulin stability in horses at risk of insulin dysregulation.. J Vet Intern Med 2019 Nov;33(6):2746-2751.
    doi: 10.1111/jvim.15629pubmed: 31617618google scholar: lookup
  13. 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.
    doi: 10.1371/journal.pone.0220203pubmed: 31339945google scholar: lookup
  14. 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.
    doi: 10.1111/jvim.15423pubmed: 30724412google scholar: lookup
  15. Fitzgerald DM, Walsh DM, Sillence MN, Pollitt CC, de Laat MA. Insulin and incretin responses to grazing in insulin-dysregulated and healthy ponies.. J Vet Intern Med 2019 Jan;33(1):225-232.
    doi: 10.1111/jvim.15363pubmed: 30506731google scholar: lookup
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