The Journal of small animal practice · 2026
Validation of an ELISA assay for measurement of the metabolite of serotonin, 5-hydroxyindole acetic acid (5-HIAA), in canine urine.
СтатьяВалидационное исследованиеНазвание и аннотация: Перевод пока не подготовлен
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Castillo D, Swallow A, Williams T, Watson P
Аннотация
OBJECTIVES: Serotonin (5-hydroxytryptophan), implicated in a number of canine diseases, has a very short half-life in the serum. Urine concentration of its breakdown product 5-hydroxyindole acetic acid after an 8 hour fast is a more reliable measure of circulating serotonin in humans. This study aimed to validate a commercially available ELISA assay to measure 5-hydroxyindole acetic acid concentrations in canine urine by comparing the analytical performance with the gold standard of liquid chromatography-tandem mass spectrometry.
METHODS: Urine was collected from 26 dogs undergoing routine diagnostic investigations at one referral centre and rapidly processed and stored prior to testing for 5-hydroxyindole acetic acid using the two methods. Deming regression and Bland-Altman analyses were used to compare the results between the two methods.
RESULTS: The ELISA demonstrated acceptable precision and repeatability (coefficient of variation <20%). There was good agreement between the two methods (bias 0.92 μmol/L; 95% limits of agreement -6.44 to 8.29 μmol/L), although the ELISA was not tested at values close to the upper-end of the claimed analytical measurement range limit. The ELISA was likely to be very reliable at low concentrations but may exceed acceptable error limits at high concentrations.
CLINICAL SIGNIFICANCE: A commercially available ELISA was validated to measure urine 5-hydroxyindole acetic acid. This is a less invasive method than blood sampling for serotonin and should give a more reliable indication of long-term serum serotonin concentrations. More studies of normal and diseased dogs are needed to confirm these findings before applying them in a clinical setting.
METHODS: Urine was collected from 26 dogs undergoing routine diagnostic investigations at one referral centre and rapidly processed and stored prior to testing for 5-hydroxyindole acetic acid using the two methods. Deming regression and Bland-Altman analyses were used to compare the results between the two methods.
RESULTS: The ELISA demonstrated acceptable precision and repeatability (coefficient of variation <20%). There was good agreement between the two methods (bias 0.92 μmol/L; 95% limits of agreement -6.44 to 8.29 μmol/L), although the ELISA was not tested at values close to the upper-end of the claimed analytical measurement range limit. The ELISA was likely to be very reliable at low concentrations but may exceed acceptable error limits at high concentrations.
CLINICAL SIGNIFICANCE: A commercially available ELISA was validated to measure urine 5-hydroxyindole acetic acid. This is a less invasive method than blood sampling for serotonin and should give a more reliable indication of long-term serum serotonin concentrations. More studies of normal and diseased dogs are needed to confirm these findings before applying them in a clinical setting.