Validation of a paraoxon-based method for measurement of paraoxonase (PON-1) activity and establishment of RIs in horses.
Abstract: Paraoxonase-1 (PON-1) is an antioxidant compound that is considered a negative acute phase protein. No information on the analytic performance of the paraoxon method for measuring PON-1 in horse serum is available. Objective: The aim of this study was to validate a paraoxon-based method to measure PON-1 in horses and to establish RIs in healthy horses and foals. Methods: Horses and foals classified as healthy after physical examination and routine biochemistry were used in the study. Serum PON-1 activity was measured with an automated spectrophotometer and an enzymatic method validated in other species. After the analytic validation (precision, accuracy, interference studies), RIs were determined using the Reference Value Advisor software. The possible sex-, age-, and breed-related differences were statistically investigated. Results: The healthy study population included 120 horses and 55 foals. The paraoxon-based method was precise (CVs < 4.0%) and accurate (P < .001 in linearity under dilution and spike-recovery testing) but was affected by interference from mild bilirubinemia, severe lipemia, and hemoglobinemia. The RIs recorded in the whole population were 38.1-80.8 U/mL. According to the Harris and Boyd test, it would be advisable to use separate RIs only for adult females and for Warmblood and Trotter adults. Conclusions: This study demonstrated that the analytic performances of the paraoxon-based method for measurement of PON-1 in horses are acceptable. The PON-1 activity is lower in horses than in other domestic species. These results may provide a basis for further studies designed to establish whether healthy and sick horses can be correctly classified by using the PON-1 assay.
© 2018 American Society for Veterinary Clinical Pathology.
Publication Date: 2018-03-27 PubMed ID: 29575140DOI: 10.1111/vcp.12562Google Scholar: Lookup
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- Journal Article
- Validation Study
Summary
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The research validates an experimental method using paraoxon to measure the activity of an enzyme called paraoxonase-1 (PON-1) in horses. This validation was also purposed towards setting reference intervals (RIs) for this enzyme in healthy horses and foals.
Context and Aim
- Paraoxonase-1 (PON-1) is an enzyme known for its antioxidant properties and is considered as a ‘negative acute phase protein’.
- However, there had not been any existing information on the performance of a method using paraoxon for evaluating PON-1 levels in horse serum.
- The research was thus aimed at validating such a method for horses, and also to establish standard RIs in healthy horses and foals.
Methodology
- The study was carried out on horses and foals deemed healthy after a physical exam and routine biochemistry.
- The activity of PON-1 in the serum was tracked using an automated spectrophotometer and an enzymatic method that had been validated in other species.
- After analytical validation involving precision, accuracy, and interference studies, the RIs were ascertained using the Reference Value Advisor software.
- Any differences in the results due to sex, age, or breed were separately noted and statistically analysed.
Results
- 120 horses and 55 foals were determined to be healthy for the study.
- The method using paraoxon was found to have acceptable precision and accuracy, although it experienced interference from conditions like mild bilirubinemia, severe lipemia, and hemoglobinemia.
- The RIs noted in the entire population ranged from 38.1 to 80.8 U/mL.
- The Harris and Boyd test indicated that separate RIs should be implemented only for adult females, Warmblood adults, and Trotter adults.
Conclusions
- This study shows that the analytical performances of using the paraoxon-based method for measuring PON-1 levels in horses are satisfactory.
- The levels of PON-1 activity are lower in horses than in other domestic species.
- The findings call for further studies to understand whether healthy and sick horses can be correctly classified using this PON-1 assay.
Cite This Article
APA
Ruggerone B, Bonelli F, Nocera I, Paltrinieri S, Giordano A, Sgorbini M.
(2018).
Validation of a paraoxon-based method for measurement of paraoxonase (PON-1) activity and establishment of RIs in horses.
Vet Clin Pathol, 47(1), 69-77.
https://doi.org/10.1111/vcp.12562 Publication
Researcher Affiliations
- Department of Veterinary Medicine, University of Milan, Milan, Italy.
- Veterinary Teaching Hospital, University of Milan, Lodi, Italy.
- Department of Veterinary Sciences, University of Pisa, San Piero a Grado, Italy.
- Department of Veterinary Sciences, University of Pisa, San Piero a Grado, Italy.
- Department of Veterinary Medicine, University of Milan, Milan, Italy.
- Veterinary Teaching Hospital, University of Milan, Lodi, Italy.
- Department of Veterinary Medicine, University of Milan, Milan, Italy.
- Veterinary Teaching Hospital, University of Milan, Lodi, Italy.
- Department of Veterinary Sciences, University of Pisa, San Piero a Grado, Italy.
MeSH Terms
- Animals
- Aryldialkylphosphatase / blood
- Blood Chemical Analysis / methods
- Blood Chemical Analysis / veterinary
- Female
- Horses / blood
- Male
- Reference Values
Citations
This article has been cited 5 times.- Rossi G, Richardson A, Jamaludin H, Secombe C. Preanalytical variables affecting the measurement of serum paraoxonase-1 activity in horses. J Vet Diagn Invest 2021 Jan;33(1):59-66.
- Scavone D, Sgorbini M, Borges AS, Oliveira-Filho JP, Vitale V, Paltrinieri S. Serial measurements of Paraoxonase-1 (PON-1) activity in horses with experimentally induced endotoxemia. BMC Vet Res 2020 Nov 4;16(1):422.
- Rossi G, Meazzi S, Giordano A, Paltrinieri S. Serum paraoxonase 1 activity in cats: analytical validation, reference intervals, and correlation with serum amyloid A and alpha-1-acid glycoprotein. J Vet Diagn Invest 2020 Nov;32(6):844-855.
- Ruggerone B, Paltrinieri S, Giordano A, Scavone D, Nocera I, Rinnovati R, Spadari A, Scacco L, Pratelli P, Sgorbini M. Paraoxonase-1 activity evaluation as a diagnostic and prognostic marker in horses and foals. J Vet Intern Med 2020 Mar;34(2):949-954.
- Baldin M, Gelain ME, Marolato G, Bedin S, Berlanda M, Zanetti M, Torrigiani F, Giordano A, Moretti P, Scavone D, Bonsembiante F. Beyond Individual Acute Phase Protein Assessments: Introducing the Acute Phase Index (API) as a Prognostic Indicator in Dogs with Malignant Neoplasia. Vet Sci 2025 Jun 1;12(6).
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