Validation and reference range determination of Aldosterone ELISA in clinically healthy dogs

Aldosterone plays a key role in maintaining electrolyte and fluid balance through its effects on renal sodium retention and potassium excretion. Despite its clinical relevance, validated assays for measuring aldosterone concentrations in dogs are limited.

This study aimed to validate a commercially available enzyme-linked immunosorbent assay (ELISA) for measuring aldosterone in canine serum samples and to establish a reference range in clinically healthy dogs.

Serum samples from 75 clinically inapparent dogs were analyzed. The dogs ranged in age from 0,5 to 12,2 years with a median age of 3,3 years. A wide variety of breeds were represented in the study population. Aldosterone concentrations were determined using an ELISA kit (ADI-900-173, Enzo Life Sciences Inc., Lausen, Switzerland) according to manufacturer’s instructions. Validation procedures included assessment of intra-assay and inter-assay precision, as well as linearity via dilution series. The intra-assay coefficient of variation (CV) ranged from 4.4% to 6.6%, while the inter-assay CV ranged from 10.8% to 18.0%. Total T4 concentration (TT4), thyroid stimulating hormone (TSH) and thyroglobuline antibodies (TgAA) were also determined. Dogs with positive TgAA, low TT4 levels and elevated TSH levels were excluded from the study (n=5), leaving a total of 70 dogs for the calculation of the aldosterone reference interval. Measurements of serum aldosterone concentrations showed wide variability with an upper reference limit (95% quantile) defined as<132,05 pg/ml (range:<32.0 to 368.4 pg/ml).

The successfully validated ELISA can be reliably used for aldosterone measurement in dogs and a reference limit was established based on the study population.

Publication History

Article published online:
11 February 2026