Research article - Peer-reviewed, 2022
Quantification of normetanephrine in canine urine using ELISA: evaluation of factors affecting results
Hoglund, Katja; Palmqvist, Hanna; Ringmark, Sara; Svensson, AnnaAbstract
Catecholamine release increases in dogs with pheochromocytomas and in situations of stress. Although plasma catecholamines degrade rapidly, their metabolites, normetanephrine (NME) and metanephrine (ME), are stable in acidified urine. Our aim was to verify a human urine ELISA kit for the quantification of NME and ME in canine urine and to determine the effects on metabolite stability of sampling time (morning or midday) and day (ordinary or day spent in a clinic). We analyzed 179 urine samples from 17 healthy dogs. For NME, the mean intra-assay CV was 6.0% for all samples and 4.3% for the canine control; inter-assay CVs were 3.3, 3.8, and 12% for high and low concentration human urine positive controls supplied in the ELISA kit and a positive canine control, respectively; spike-recovery was 90-101%. For ME, mean intra-assay CV was 6.5% for samples and 9.0% for the canine control; inter-assay CVs were 12.7, 7.2, and 22.5% for high and low concentration human urine positive controls supplied in the ELISA kit and a positive canine control, respectively; spike-recovery was 85-89%. Dilution recovery was unsatisfactory for both metabolites. Based on our verification results, NME was selected for remaining analyses. We found no effect on NME concentrations of acidification or room temperature storage for up to 24 h. The NME:creatinine ratio was higher after the first of 3 clinic days compared to the same morning (111.2 +/- 5.5 vs. 82.9 +/- 5.3; p < 0.0001), but not on the other days. NME verification results were generally superior to ME. Dilution studies were unsatisfactory for both metabolites. Given that NME was stable without acidification at room temperature, urine samples can be collected at home. The clinic environment can cause higher NME:creatinine ratios, especially in unaccustomed dogs.Keywords
acidification; catecholamine; dogs; metanephrine; metanephrines; normetanephrine; sample storagePublished in
Journal of Veterinary Diagnostic Investigation2022, volume: 34, number: 1, article number: 10406387211052984
Publisher: SAGE PUBLICATIONS INC
Authors' information
Swedish University of Agricultural Sciences, Department of Anatomy, Physiology and Biochemistry (AFB)
Swedish University of Agricultural Sciences, Department of Anatomy, Physiology and Biochemistry (AFB)
Swedish University of Agricultural Sciences, Department of Animal Nutrition and Management
Swedish University of Agricultural Sciences, Department of Clinical Sciences
UKÄ Subject classification
Pathobiology
Publication Identifiers
DOI: https://doi.org/10.1177/10406387211052984
URI (permanent link to this page)
https://res.slu.se/id/publ/114300