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BMC veterinary research2016; 12(1); 104; doi: 10.1186/s12917-016-0737-1

Analytical validation and establishment of reference intervals for a ‘high-sensitivity’ cardiac troponin-T assay in horses.

Abstract: Cardiac troponin-I assays have been validated in horses.'High-sensitivity' cardiac troponin assays are now the standard in human cardiology. Objective: Appropriately validate the'high-sensitivity' cardiac Troponin-T (hscTnT) assay for clinical use in horses, establish reference intervals, determine the biological variation, and demonstrate assay utility in selected clinical cases. Methods: Analytical validation of the Roche hscTnT assay included within- and between-run precision, linear dose response, limit of quantitation (LoQ), stability, and comparison with cTn-I (iSTAT). Reference intervals and biological variation were determined using adult, healthy, Non-Competition Horses (N = 125) and Racing-Thoroughbreds (N = 178). HscTnT levels were measured in two horses with cardiac pathology. Results: The hscTnT demonstrates acceptable within-run (L1 = 6.5 ng/L, CV 14.9 %, L2 = 10.1 ng/L, CV 8.7 %, L3 = 15.3 ng/L, CV 5.4 %) and between-run precision (L1 = 12.2 ng/L, CV 8.4 %, L2 = 57.0 ng/L, CV 8.4 %, L3 = 256.0 ng/L, CV 9.0 %). The assay was linear from 3 to 391 ng/L. The LoQ was validated at 3 ng/L. Samples demonstrated insignificant decay over freeze-thaw cycle. Comparison with cTnI assay showed excellent correlation (range: 8.0-3535.0 ng/L, R(2) = 0.9996). Reference intervals: The upper 95(th) and 99(th) percentile of the hscTnT population distribution were 6.8 and 16.2 ng/L in Non-Competition Horses, and 14.0 and 23.2 ng/L in Racing-Thoroughbreds. Between-breed, diurnal effect, and between-day variation was below LoQ. Two clinical cases with presumed cardiac pathology had hscTnT levels of 220.9 ng/L and 5723.0 ng/L. Conclusions: This benchmark study is the first to comply with CLSI guidelines, thus further establishing the performance characteristics of the hscTnT assay, and reference intervals in healthy horses. Two clinical cases demonstrated further the clinical utility of the assay.
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Publication Date: 2016-06-13 PubMed ID: 27296016PubMed Central: PMC4906598DOI: 10.1186/s12917-016-0737-1Google Scholar: Lookup
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Summary

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This research study validates the use of ‘high-sensitivity’ cardiac Troponin-T (hscTnT) assays for clinical use in horses. The study conducted the necessary analytical validations, provided reference intervals, examined the biological variation, and demonstrated the clinical utility of the test in detecting cardiac pathology in horses.

Methodology

  • The study used the Roche hscTnT assay and evaluated its within- and between-run precision, linear dose response, limit of quantitation (LoQ), and stability. The researchers compared it with a Cardiac Troponin-I (cTnI) assay.
  • The reference intervals and biological variation data were determined using two groups of horses: Non-Competition Horses (125 horses) and Racing-Thoroughbreds (178 horses).
  • The hscTnT levels were also measured in two horses with cardiac pathology to demonstrate the clinical utility of the test.

Results

  • The hscTnT assay delivered acceptable within-run and between-run precision results, indicating a reliable performance.
  • The assay was found to be linear in its response, meaning that the change in the output, or the concentration of troponin, is directly proportional to the change in the input.
  • The LoQ, or the lowest quantity of a substance that can be distinguished from the absence of that substance, was validated at 3 ng/L. The stable performance of the assay over a freeze-thaw cycle also demonstrated its robustness.
  • The comparison with the cTnI assay showed an excellent correlation, indicating that the hscTnT can be used in place of the cTnI assay.
  • Different values for upper 95th and 99th percentiles of the hscTnT population distribution were identified for Non-Competition Horses and Racing-Thoroughbreds. This indicates that reference values could change depending on the type of horse, and thus should be considered while interpreting results.
  • The impact of factors such as the breed of the horse, time of day, and day-to-day variation was below LoQ.
  • The application of the assay on two clinical cases with cardiac pathology validated the clinical utility of the assay.

Conclusion

  • The research concludes by affirming the clinical use of hscTnT assay in horses. This validation followed CLSI (Clinical Laboratory Standards Institute) guidelines, providing a robust and standardized methodology.
  • It also established reference intervals to assist practitioners in interpreting the results of the hscTnT assay. These reference values are especially crucial for recognizing out of range results, which might indicate the presence of cardiac pathology.
  • The research exemplifies the utility of the assay in clinical cases, thus demonstrating its practicality for cardiac health screening in horses.

Cite This Article

APA
Shields E, Seiden-Long I, Massie S, Passante S, Leguillette R. (2016). Analytical validation and establishment of reference intervals for a ‘high-sensitivity’ cardiac troponin-T assay in horses. BMC Vet Res, 12(1), 104. https://doi.org/10.1186/s12917-016-0737-1

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 12
Issue: 1
Pages: 104
PII: 104

Researcher Affiliations

Shields, E
  • University of Calgary Faculty of Veterinary Medicine (UCVM), 3330 Hospital Dr. NW, Calgary, T2N 4 N1, AB, Canada.
Seiden-Long, I
  • Foothills Medical Centre, University of Calgary Faculty of Medicine and Calgary Lab Services (CLS), Room C618B, 1403-29th St. NW, Calgary, T2N 2 T9, AB, Canada.
Massie, S
  • University of Calgary Faculty of Veterinary Medicine (UCVM), 3330 Hospital Dr. NW, Calgary, T2N 4 N1, AB, Canada.
Passante, S
  • University of Calgary Faculty of Veterinary Medicine (UCVM), 3330 Hospital Dr. NW, Calgary, T2N 4 N1, AB, Canada.
Leguillette, R
  • University of Calgary Faculty of Veterinary Medicine (UCVM), 3330 Hospital Dr. NW, Calgary, T2N 4 N1, AB, Canada. rleguill@ucalgary.ca.

MeSH Terms

  • Animals
  • Female
  • Horses / blood
  • Humans
  • Immunoassay / methods
  • Immunoassay / veterinary
  • Luminescent Measurements / veterinary
  • Male
  • Reference Values
  • Sensitivity and Specificity
  • Troponin T / analysis

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Citations

This article has been cited 4 times.
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