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Home / Equine Research Bank / Animals (Basel) / Validation of a Commercial ELISA Kit for Non-Invasive Measur...
Animals : an open access journal from MDPI2024; 14(19); 2831; doi: 10.3390/ani14192831

Validation of a Commercial ELISA Kit for Non-Invasive Measurement of Biologically Relevant Changes in Equine Cortisol Concentrations.

Abstract: The measurement of fecal cortisol/corticosterone metabolites (FCMs) is often used to quantify the stress response. The sampling method is relatively non-invasive, reduces concern for elevation of cortisol from the sampling method, and has been shown to measure cortisol more consistently without the daily diurnal rhythm observed in blood. Commercial ELISA (enzyme-linked immunoassay) kits offer benefits over previously validated immunoassay methods but lack validation. The objective of this study was to evaluate a commercial ELISA kit (Arbor Assays DetectX Cortisol ELISA kit, K003-H1, Ann Arbor, MI, USA) and provide analytical and biologic validation of equine fecal and plasma samples. Horses (4 male, 4 female, mean ± SD: 4 ± 5 yr) were transported for 15 min with limited physical and visual contact via a livestock trailer. Blood and fecal samples were collected pre- and post-transportation. Parallelism, accuracy, and precision tests were used to analytically validate this kit. Data were analyzed using PROC MIXED in SAS 9.4. Plasma cortisol concentrations increased in response to trailering (254.5 ± 26.4 nmol/L, 0 min post-transportation) compared to pre-transportation (142.8 ± 26.4 nmol/L). FCM concentrations increased 24 h post-trailering (10.8 ± 1.7 ng/g) when compared to pre-transportation (7.4 ± 1.7 ng/g). These data support that changes in FCMs can be observed 24 h post-stressor. In conclusion, the Arbor Assays DetectX Cortisol ELISA kit is a reliable, economic option for the measurement of biologically relevant changes in cortisol in equine plasma and FCMs.
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Publication Date: 2024-10-01 PubMed ID: 39409780PubMed Central: PMC11475127DOI: 10.3390/ani14192831Google 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.

The research article discusses the use and validation of a commercial test known as the Arbor Assays DetectX Cortisol ELISA kit, for measuring stress levels in horses by detecting changes in cortisol concentration in their feces and blood. The study finds that this kit is a reliable and cost-effective method for such measurements.

Study Objectives and Methodology

  • The primary objective of this study was to evaluate the Arbor Assays DetectX Cortisol ELISA kit for its ability to accurately and consistently measure cortisol levels in equine fecal and plasma samples. This cortisol ELISA kit is a commercial product that offers some benefits over traditional immunoassay methods, but it has not been validated before this study.
  • For the purpose of this experiment, eight horses were involved, comprising of four males and four females, with a mean age of 4 ± 5 years. The horses were transported for 15 minutes in a livestock trailer with limited physical and visual contact to generate a stress response.
  • Blood and fecal samples were obtained from the horses both before and after transportation. The analytical validation of the ELISA kit involved executing tests for parallelism, accuracy, and precision.
  • The analytical data generated was processed using PROC MIXED in SAS 9.4.

Results and Interpretation

  • The results showed an increase in plasma cortisol concentrations in the horses as a result of the transportation stressor (254.5 ± 26.4 nmol/L measured immediately post-transportation, compared to 142.8 ± 26.4 nmol/L before transportation).
  • Fecal cortisol metabolite (FCM) concentrations also increased 24 hours after the stressor (10.8 ± 1.7 ng/g post-transportation, compared to 7.4 ± 1.7 ng/g pre-transportation). This signifies that changes in FCMs as a stress response can be observed as late as 24 hours after the stressor.

Conclusions and Implications

  • In conclusion, the Arbor Assays DetectX Cortisol ELISA kit is validated, indicating that it is a reliable, cost-effective alternative for measuring biologically relevant changes in cortisol levels in equine plasma and fecal samples.
  • The ability to accurately measure stress levels in horses through non-invasive means such as this can have significant applications in equine health care and management. It allows for the detection of the stress response without causing additional stress to the animal from invasive procedures.

Cite This Article

APA
Share ER, Mastellar SL, Suagee-Bedore JK, Eastridge ML. (2024). Validation of a Commercial ELISA Kit for Non-Invasive Measurement of Biologically Relevant Changes in Equine Cortisol Concentrations. Animals (Basel), 14(19), 2831. https://doi.org/10.3390/ani14192831

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 14
Issue: 19
PII: 2831

Researcher Affiliations

Share, Elizabeth R
  • Department of Animal Sciences, The Ohio State University, Columbus, OH 43210, USA.
Mastellar, Sara L
  • Agricultural Technical Institute, The Ohio State University, Wooster, OH 44691, USA.
Suagee-Bedore, Jessica K
  • Department of Animal and Poultry Sciences, Virginia Tech University, Blacksburg, VA 24061, USA.
Eastridge, Maurice L
  • Department of Animal Sciences, The Ohio State University, Columbus, OH 43210, USA.

Conflict of Interest Statement

The authors declare no conflicts of interest.

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Citations

This article has been cited 2 times.
  1. Arshad A, Reif AH, Cavalleri JV, Desvars-Larrive A. Zoonotic pathogens in equids in Central Europe: a systematic review. BMC Vet Res 2025 Jul 8;21(1):451.
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  2. Nowak AC, Macho-Maschler S, Biermann NM, Palme R, Dengler F. Investigating the interplay of stressors and health in horses through fecal cortisol metabolite analysis. Front Vet Sci 2025;12:1545577.
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