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Quantification of normetanephrine in canine urine using ELISA: evaluation of factors affecting results
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 quantificat...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
SAGE Publications
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8689017/ https://www.ncbi.nlm.nih.gov/pubmed/34697963 http://dx.doi.org/10.1177/10406387211052984 |
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author | Höglund, Katja Palmqvist, Hanna Ringmark, Sara Svensson, Anna |
author_facet | Höglund, Katja Palmqvist, Hanna Ringmark, Sara Svensson, Anna |
author_sort | Höglund, Katja |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8689017 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | SAGE Publications |
record_format | MEDLINE/PubMed |
spelling | pubmed-86890172021-12-22 Quantification of normetanephrine in canine urine using ELISA: evaluation of factors affecting results Höglund, Katja Palmqvist, Hanna Ringmark, Sara Svensson, Anna J Vet Diagn Invest Full Scientific Reports 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. SAGE Publications 2021-10-26 2022-01 /pmc/articles/PMC8689017/ /pubmed/34697963 http://dx.doi.org/10.1177/10406387211052984 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution 4.0 License (https://creativecommons.org/licenses/by/4.0/) which permits any use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage). |
spellingShingle | Full Scientific Reports Höglund, Katja Palmqvist, Hanna Ringmark, Sara Svensson, Anna Quantification of normetanephrine in canine urine using ELISA: evaluation of factors affecting results |
title | Quantification of normetanephrine in canine urine using ELISA: evaluation of factors affecting results |
title_full | Quantification of normetanephrine in canine urine using ELISA: evaluation of factors affecting results |
title_fullStr | Quantification of normetanephrine in canine urine using ELISA: evaluation of factors affecting results |
title_full_unstemmed | Quantification of normetanephrine in canine urine using ELISA: evaluation of factors affecting results |
title_short | Quantification of normetanephrine in canine urine using ELISA: evaluation of factors affecting results |
title_sort | quantification of normetanephrine in canine urine using elisa: evaluation of factors affecting results |
topic | Full Scientific Reports |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8689017/ https://www.ncbi.nlm.nih.gov/pubmed/34697963 http://dx.doi.org/10.1177/10406387211052984 |
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