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

J Vet Intern Med. 2014 Sep-Oct;28(5):1433-41. doi: 10.1111/jvim.12399. Epub 2014 Jul 16.

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.

Keywords: Hospital stress; Influence; Reference interval; UCCR; Variation.

MeSH terms

  • Adrenal Cortex Hormones / urine*
  • Animals
  • Creatinine / urine
  • Cushing Syndrome / urine
  • Cushing Syndrome / veterinary*
  • Dog Diseases / urine*
  • Dogs / urine*
  • Female
  • Gas Chromatography-Mass Spectrometry / veterinary*
  • Immunoassay / veterinary*
  • Luminescent Measurements / veterinary
  • Male
  • Prospective Studies
  • Radioimmunoassay / veterinary

Substances

  • Adrenal Cortex Hormones
  • Creatinine