Correcting for enzyme immunoassay changes in long term monitoring studies

Abstract

Enzyme immunoassays (EIAs) are a common tool for measuring steroid hormones in wildlife due to their low cost, commercial availability, and rapid results. Testing technologies improve continuously, sometimes requiring changes in protocols or crucial assay components. Antibody replacement between EIA kits can cause differences in EIA sensitivity, which can hinder monitoring hormone concentration over time. The antibody in a common cortisol EIA kit used for long-term monitoring of stress in wildlife was replaced in 2014, causing differences in cross reactivity and standard curve concentrations. Therefore, the objective of this study was to develop a method to standardize results following changes in EIA sensitivity. We validated this method using cortisol concentrations measured in the hair of brown bears ( Ursus arctos ). We used a simple linear regression to model the relationship between cortisol concentrations using kit 1 and kit 2. We found a linear relationship between the two kits (R 2 = 0.85) and used the regression equation (kit2 = (0.98 × kit1) + 1.65) to predict cortisol concentrations in re-measured samples. Mean predicted percent error was 16% and 72% of samples had a predicted percent error <20%, suggesting that this method is well-suited for correcting changes in EIA sensitivity.

Hormonal and Neurological Aspects of Dog Walking for Dog Owners and Pet Dogs

“…We thawed the frozen cotton swabs containing saliva in 10 mL separation tubes, centrifuged them at 3500 rpm for 15 min at 4 °C, and then used the separated fluids as samples. We measured hormone levels in saliva with the enzyme-linked immunosorbent assay (ELISA) kits, Enzo Biochem (Enzo Life Sciences Inc., Farmingdale, NY, USA), [ 23 ] and Oxford Biomedical Research (Rochester Hills, MI, USA) [ 24 ] for oxytocin and cortisol, respectively. These kits have reduced non-specific binding, and therefore are more accurate.…”