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Urinary corticoid concentrations measured by 5 different immunoassays and gas chromatography-mass spectrometry in healthy dogs and dogs with hypercortisolism at home and in the hospital


Galeandro, L; Sieber-Ruckstuhl, N S; Riond, B; Hartnack, S; Hofmann-Lehmann, R; Reusch, C E; Boretti, F S (2014). Urinary corticoid concentrations measured by 5 different immunoassays and gas chromatography-mass spectrometry in healthy dogs and dogs with hypercortisolism at home and in the hospital. Journal of Veterinary Internal Medicine, 28(5):1433-1441.

Abstract

BACKGROUND Determination of the urinary corticoid-to-creatinine ratio (UCCR) is an important screening test in the diagnosis of hypercortisolism (HC). However, urinary cortisol metabolites interfere with cortisol measurement in immunoassays, leading to decreased specificity. Gas chromatography-mass spectrometry (GC-MS) is considered the gold standard for steroid hormone analysis, because it provides a high level of selectivity and accuracy. OBJECTIVES To prospectively compare the UCCR of healthy dogs and dogs with HC determined by 5 different immunoassays and by GC-MS and to evaluate the influence of veterinary care on UCCR. ANIMALS Twenty healthy dogs; 18 dogs with HC. METHODS Urine was collected in the hospital and again after 6 days at home. Three chemiluminescence immunoassays (Access 2, Beckmann; Immulite 2000, DPC Siemens, with and without trichloromethane extraction) and 2 RIAs (Utrecht in house; Access Beckmann) were used. GC-MS analyses were performed with Agilent 6890N/5973N. Urinary corticoid concentrations were related to urinary creatinine concentrations. RESULTS Immunoassay results were significantly higher compared to GC-MS results. Evaluation of bias plots and clinical assessment made on the basis of the assay results of each dog indicated substantial disagreement among the assays. Sensitivity varied from 37.5 to 75% and with selected assays was lower in samples from day 6 compared to day 0. GC-MS was not superior to the immunoassays in discriminating healthy from HC dogs. CONCLUSIONS AND CLINICAL IMPORTANCE Considerable variation must be anticipated comparing different urinary cortisol assays. Establishing an assay- and laboratory-specific reference range is critical when using UCCR.

BACKGROUND Determination of the urinary corticoid-to-creatinine ratio (UCCR) is an important screening test in the diagnosis of hypercortisolism (HC). However, urinary cortisol metabolites interfere with cortisol measurement in immunoassays, leading to decreased specificity. Gas chromatography-mass spectrometry (GC-MS) is considered the gold standard for steroid hormone analysis, because it provides a high level of selectivity and accuracy. OBJECTIVES To prospectively compare the UCCR of healthy dogs and dogs with HC determined by 5 different immunoassays and by GC-MS and to evaluate the influence of veterinary care on UCCR. ANIMALS Twenty healthy dogs; 18 dogs with HC. METHODS Urine was collected in the hospital and again after 6 days at home. Three chemiluminescence immunoassays (Access 2, Beckmann; Immulite 2000, DPC Siemens, with and without trichloromethane extraction) and 2 RIAs (Utrecht in house; Access Beckmann) were used. GC-MS analyses were performed with Agilent 6890N/5973N. Urinary corticoid concentrations were related to urinary creatinine concentrations. RESULTS Immunoassay results were significantly higher compared to GC-MS results. Evaluation of bias plots and clinical assessment made on the basis of the assay results of each dog indicated substantial disagreement among the assays. Sensitivity varied from 37.5 to 75% and with selected assays was lower in samples from day 6 compared to day 0. GC-MS was not superior to the immunoassays in discriminating healthy from HC dogs. CONCLUSIONS AND CLINICAL IMPORTANCE Considerable variation must be anticipated comparing different urinary cortisol assays. Establishing an assay- and laboratory-specific reference range is critical when using UCCR.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Chair in Veterinary Epidemiology
05 Vetsuisse Faculty > Veterinary Clinic > Department of Small Animals
05 Vetsuisse Faculty > Veterinary Clinic > Department of Farm Animals
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Language:English
Date:2014
Deposited On:30 Oct 2014 09:55
Last Modified:05 Apr 2016 18:26
Publisher:Wiley-Blackwell Publishing, Inc.
ISSN:0891-6640
Publisher DOI:https://doi.org/10.1111/jvim.12399
PubMed ID:25040917

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