Proceeding of Veterinary and Animal Science Days 2018, 6th- 8th June, Milan, Italy 

HAF © 2013 

Vol. V, No. 1s  ISSN: 2283-3927 

   

 

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Aldosterone is a corticosteroid hormone that plays a pivotal role in homeostatic regulation of 

water and salt reabsorption, blood volume and pressure. Aldosterone levels tent to rise in 

humans in hypertension, chronic and acute congestive heart failure (CHF); detrimental effects 

are opposed by drugs like ACE inhibitors and anti-mineralocorticoid. Aldosterone has a pulsatile 

secretion, so measurement in serum is less indicative than in urine, where concentration can be 

indexed to creatinine ratio for estimation of the 24-h aldosterone excretion. 

Few studies have evaluated aldosterone in canine urine patients, and none by ELISA. Aim of the 

study was to evaluate a commercial ELISA kit for measuring aldosterone in dog’s urine. 

Urine was collected by free catch from four dogs. Two were healthy, one was affected by CHF 

and prescribed anti-mineralocorticoid daily, one was affected by chronic kidney disease (CKD). 

Urine was centrifuged (1250g/5 min) and supernatant frozen (-20°C). Aldosterone was measured 

by a competitive ELISA previously validated for dogs. Twenty-four hours acid hydrolysis was 

performed on urinary samples before assay. 

The ELISA standard curve in a semi-log plot was linear between 2.5 and 3.9 ng/mL. Spike-and-

recovery, linearity-of-dilution and parallelism experiments showed accuracy in measuring 

aldosterone in dog urine samples (Syme et al., 2007). Concentrations of urine aldosterone are 

reported in Table 1. The intra-assay coefficient of variation showed good reproducibility of the 

assay. 

Urinary samples are easy to collect, and the ELISA used in this preliminary study seems promising 

in determining aldosterone in dog urine. Its levels can be of great diagnostic and prognostic 

value for dogs affected by acute and chronic CHF, in order to assess the best therapeutic 

strategy (Gardner et al., 2007). This preliminary analysis will be followed by further studies in 

patients affected by acute and chronic CHF. 

 

 

 

 

Keywords 

Competitive ELISA, Myxomatous 

mitral valve disease, ACE-

inhibitors, Angiotensin II 

receptor blockers, Heart failure.  

 
CORRESPONDING AUTHOR 

Alice Savarese 

alice.savarese@unimi.it 

 
JOURNAL HOME PAGE 

riviste.unimi.it/index.php/haf 

Preliminary evaluation of an ELISA kit 

for the detection of  aldosterone 

concentration in dog’s urine. 

A. Savarese1*, C. Locatelli1, V. Borromeo1, A. Berrini 1, A. 

Galizzi1, M. Crudo1, P.G. Brambilla1 

 

1 Department of Veterinary Medicine, University of Milan, Via Celoria 10, 
20133 Milan, Italy. 

 

 

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Proceeding of Veterinary and Animal Science Days 2018, 6th- 8th June, Milan, Italy 74 

HAF © 2013 

Vol. V, No. 1s  ISSN: 2283-3927 

 

 

 

PATIENT ALDOSTERONE CONCENTRATIONS (ng/mL) 

Healthy 1 2.593 ± 0.890 (CV 14.9%) 

Healthy 2 2.737 ± 0.39 (CV 14.3%) 

CHF 3.620 ± 0.44 (CV 12.2%) 

CKD 2.399 ± 0.42 (CV 17.5%) 

 

 

References 

Syme, H.M., Fletcher, M.G.R., Bailey, S.R., Elliott, J., 2007. Measurement of aldosterone in feline, canine and human 

urine. Journal of Small Animal Practice. 48, 202–208. 

Gardner S.Y., Atkins C.E., Rausch W.P., DeFrancesco, T.C., Chandler D.W., Keene B.W. 2007. Estimation of 24-h 

aldosterone secretion in the dog using the urine aldosterone: Creatinine ratio. Journal of Veterinary Cardiology 

9, 1 - 7.  

 

 

Table 1: Concentrations of urine aldosterone measured in four dogs. Values are 

mean ± SD of three determinations. In parenthesis the intra-assay coefficient of 

variation (CV%). 

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