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Analytic validation of a paraoxon-based method to measure the activity of paraoxonase-1 in feline blood
Journal article   Peer reviewed

Analytic validation of a paraoxon-based method to measure the activity of paraoxonase-1 in feline blood

G. Rossi, A. Giordano, E. Costarelli, P. Moretti and S. Paltrinieri
Veterinary Clinical Pathology, Vol.43(4), pp.E8-E21
2014
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Abstract

The activity of the antioxidant enzyme paraoxonase (PON1) has been shown to decrease during inflammation in many species but not in cats. The aim of this study was to validate an automated method, based on the use of paraoxon, to measure PON1 activity in feline serum. Intra-assay imprecision was determined in pooled feline serum samples and accuracy was indirectly measured using linearity under dilution (LUD) and spiking-recovery tests. In order to preliminarily assess whether PON-1 activity decreases in cats with inflammation, 349 sera were analyzed with low (< 0.5 mg/mL, n = 111), medium (0.5–1.5 mg/mL, n = 113) or high (> 1.5 mg/mL, n = 125) concentration of the acute phase protein alpha-1-acid glycoprotein (AGP). Intra-assay coefficient of variation (CV) was 3.7%, 1.0%, 1.9% for samples with normal (117.1 U/L), low (59.5 U/L) and very low (20.6 U/L) PON1 activity, respectively. Both the LUD test and the spike-recovery test fit the linear model. PON-1 activity was significantly higher (P < .001) in cats with low AGP concentration (83.0 31.0 U/L; median: 82.5 U/L) than in cats with medium (64.0 31.4 U/L; 60.1 U/L) or high (43.6 30.0 U/L; 39.1 U/L) AGP concentration. The automated paraoxon-based method tested in this study has an acceptable intra-assay precision and accuracy to be used in routine practice and may differentiate samples characterized by inflammation according to the concentration of AGP. However, cats with low, medium and high AGP had overlapping PON1 activity, likely because the magnitude of PON-1 decrease depends on the presence of oxidative stress.

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