Analytical validation of a high-sensitivity cardiac troponin T assay in horses.
Abstract: Although cardiac troponin T (cTnT) assays have been used to detect myocardial damage in horses, a cTnT assay has not been analytically validated, to our knowledge. The aims of this study were to estimate the precision of a high-sensitivity cTnT assay in horses and determine the effect of hemolysis on the measured cTnT concentration. Serum samples from horses were mixed in 3 different pools. Pool 1 consisted of samples from 3 healthy horses, pool 2 from 6 horses with heart failure or atypical myopathy, and pool 3 from 10 horses with atypical myopathy. The within- and between-run coefficients of variation were determined for each pool. Pools 2 and 3 were diluted to estimate linearity. To study the influence of sample hemolysis, serum was collected from 4 horses with a high cTnT concentration, in which hemolysis was mechanically induced. In addition, ethylenediamine tetra-acetic acid blood tubes were collected from 3 other horses, from which hemolysate was prepared and added to plasma at different concentrations. The within- and between-run coefficients of variation of all pools were <10%, and a good linearity was found. Three out of 4 hemolyzed serum samples had a decreased serum cTnT concentration. Plasma samples with a high hemolysis index showed a negative interference, resulting in a lower cTnT concentration. Results of the high-sensitivity cTnT assay were highly reproducible. Because samples from horses with musculoskeletal damage were included, further studies should test the possible cross-reactivity between troponin T of musculoskeletal and cardiac origin before the assay can be used in equine clinical practice.
© 2015 The Author(s).
Publication Date: 2015-07-16 PubMed ID: 26185121DOI: 10.1177/1040638715593601Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Validation Study
Summary
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The research paper discusses the analytic validation of a sensitive cardiac Troponin T (cTnT) test used to detect heart damage in horses. It evaluates its precision and the impact of sample hemolysis or blood cell breakdown on its results.
Research Aims and Methods
- The researchers conducted tests to confirm the accuracy of a high-sensitivity cTnT assay in detecting heart damage in horses. They also sought to understand how hemolysis, the breakdown of red blood cells, affects the measured cTnT concentration.
- Serum samples were taken from various horses and divided into three pools. Pool 1 included samples from healthy horses, pool 2 from horses with heart failure or atypical myopathy (a muscle disease), and pool 3 from horses only with atypical myopathy.
- The team calculated the within-run and between-run coefficients of variation – measures of assay precision – for each pool. Pools 2 and 3 were also diluted to understand the linearity of the results.
- To analyze hemolysis’ effect, serum with a high concentration of cTnT was collected from different horses, and hemolysis was artificially induced.
- They also collected ethylenediamine tetra-acetic acid (EDTA) blood tubes from three other horses and conducted tests to analyze the variation in cTnT concentration when different levels of hemolysate are added to plasma.
Results and Conclusion
- The researchers found the assay to be highly reproducible, that is, it delivers consistent results across multiple samples, since all pools showed coefficients of variation less than 10%.
- The study also indicated a good linearity in diluted pools 2 and 3, suggesting the consistent performance of the assay across different concentration levels.
- Hemolysis was found to negatively interfere with the assay results. It decreased the cTnT concentration in the samples, meaning that the sample breakdown could potentially give a false-negative result.
- Despite the study’s promising results, the authors concluded that further investigation is required due to the inclusion of horses with musculoskeletal damage. This is an essential step to rule out potential cross-reactivity between the troponin T of musculoskeletal and cardiac origin before the test is widely used in veterinary practice.
Cite This Article
APA
Van Der Vekens N, van Dievoet MA, De Puydt H, Decloedt A, Ven S, De Clercq D, Deprez P, van Loon G.
(2015).
Analytical validation of a high-sensitivity cardiac troponin T assay in horses.
J Vet Diagn Invest, 27(4), 504-509.
https://doi.org/10.1177/1040638715593601 Publication
Researcher Affiliations
- Department of Large Animal Internal Medicine, Faculty of Veterinary Medicine-Ghent, Merelbeke, Belgium (Van Der Vekens, Decloedt, Ven, De Clercq, Deprez, van Loon)General Hospital Sint-Lucas, Ghent, Belgium (van Dievoet, De Puydt) nicky.vandervekens@ugent.be.
- Department of Large Animal Internal Medicine, Faculty of Veterinary Medicine-Ghent, Merelbeke, Belgium (Van Der Vekens, Decloedt, Ven, De Clercq, Deprez, van Loon)General Hospital Sint-Lucas, Ghent, Belgium (van Dievoet, De Puydt).
- Department of Large Animal Internal Medicine, Faculty of Veterinary Medicine-Ghent, Merelbeke, Belgium (Van Der Vekens, Decloedt, Ven, De Clercq, Deprez, van Loon)General Hospital Sint-Lucas, Ghent, Belgium (van Dievoet, De Puydt).
- Department of Large Animal Internal Medicine, Faculty of Veterinary Medicine-Ghent, Merelbeke, Belgium (Van Der Vekens, Decloedt, Ven, De Clercq, Deprez, van Loon)General Hospital Sint-Lucas, Ghent, Belgium (van Dievoet, De Puydt).
- Department of Large Animal Internal Medicine, Faculty of Veterinary Medicine-Ghent, Merelbeke, Belgium (Van Der Vekens, Decloedt, Ven, De Clercq, Deprez, van Loon)General Hospital Sint-Lucas, Ghent, Belgium (van Dievoet, De Puydt).
- Department of Large Animal Internal Medicine, Faculty of Veterinary Medicine-Ghent, Merelbeke, Belgium (Van Der Vekens, Decloedt, Ven, De Clercq, Deprez, van Loon)General Hospital Sint-Lucas, Ghent, Belgium (van Dievoet, De Puydt).
- Department of Large Animal Internal Medicine, Faculty of Veterinary Medicine-Ghent, Merelbeke, Belgium (Van Der Vekens, Decloedt, Ven, De Clercq, Deprez, van Loon)General Hospital Sint-Lucas, Ghent, Belgium (van Dievoet, De Puydt).
- Department of Large Animal Internal Medicine, Faculty of Veterinary Medicine-Ghent, Merelbeke, Belgium (Van Der Vekens, Decloedt, Ven, De Clercq, Deprez, van Loon)General Hospital Sint-Lucas, Ghent, Belgium (van Dievoet, De Puydt).
MeSH Terms
- Animals
- Biomarkers / blood
- Cardiomyopathies / blood
- Cardiomyopathies / diagnosis
- Cardiomyopathies / veterinary
- Horse Diseases / blood
- Horse Diseases / diagnosis
- Horses
- Myocardium / pathology
- Reproducibility of Results
- Troponin T / blood
Citations
This article has been cited 1 times.- Shields E, Seiden-Long I, Massie S, Passante S, Leguillette R. Analytical validation and establishment of reference intervals for a 'high-sensitivity' cardiac troponin-T assay in horses. BMC Vet Res 2016 Jun 13;12(1):104.
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