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  • Author or Editor: Luca Giori x
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Abstract

OBJECTIVE To investigate the precision of an ELISA for measurement of serum cortisol concentration (SCC) in dogs, assess agreement between this ELISA and 2 validated chemiluminescence assays (CLAs), and evaluate the clinical implications of any bias associated with this ELISA when measuring SCC in dogs.

DESIGN Evaluation study.

SAMPLE 75 stored, frozen serum samples from client-owned dogs.

PROCEDURES Enzyme-linked immunosorbent assay precision was evaluated by measuring SCC of pooled serum samples. Agreement with standard methods was evaluated with Spearman rank correlation, Passing-Bablok regression, and Bland-Altman analysis to compare SCCs obtained with the ELISA and the 2 CLAs. An error grid was used to evaluate identified bias.

RESULTS Within-laboratory coefficients of variation for pooled serum samples with low, medium, and high SCCs were 21.4%, 28.9%, and 13.0%, respectively. There was a high correlation between ELISA results (for all samples combined) and results of the 2 CLAs (CLA 1, r = 0.96; CLA 2, r = 0.97), but constant and proportional biases between the ELISA and CLAs were present at all concentrations. Clinically important disagreement between ELISA results and CLA results occurred in 16 of 63 (25%) samples, particularly with low and high SCCs.

CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that the rate of clinical disagreement between the ELISA and CLAs was sufficiently high to recommend that equivocal results obtained with the ELISA be confirmed by a reference laboratory. Further evaluation of analytic performance of the ELISA should focus on samples with very high and very low SCCs.

Restricted access
in Journal of the American Veterinary Medical Association

Abstract

Objective—To determine whether preanalytic and analytic factors affect evaluation of the urinary protein-to-creatinine (UPC) ratio in dogs.

Sample—50 canine urine samples.

Procedures—The UPC ratio was measured to assess the intra-assay imprecision (20 measurements within a single session), the influence of predilution (1:10, 1:20, and 1:100) for urine creatinine concentration measurement, and the effect of storage at room temperature (approx 20°C), 4°C, and −20°C.

Results—The coefficient of variation at room temperature determined with the 1:20 predilution was < 10.0%, with the highest coefficients of variation found in samples with a low protein concentration or low urine specific gravity. This variability could result in misclassification of samples with UPC ratios close to the thresholds defined by the International Renal Interest Society to classify dogs as nonproteinuric (0.2), borderline proteinuric (0.21 to 0.50), or proteinuric (> 0.51). A proportional bias was found in samples prediluted 1:10, compared with samples prediluted 1:20 or 1:100. At room temperature, the UPC ratio did not significantly increase after 2 and 4 hours. After 12 hours at room temperature and at 4°C, the UPC ratio significantly increased. The UPC ratio did not significantly change during 3 months of storage at −20°C.

Conclusions and Clinical Relevance—The intra-assay precision of the UPC ratio was sufficiently low to avoid misclassification of samples, except for values close to 0.2 or 0.5. The optimal predilution ratio for urine creatinine concentration measurement was 1:20. A 1:100 predilution is recommended in samples with a urine specific gravity > 1.030. The UPC ratio must be measured as soon as samples are collected. Alternatively, samples should be immediately frozen to increase their stability and minimize the risk of misclassification of proteinuria.

Full access
in American Journal of Veterinary Research