Validation of an enzyme-linked immunosorbent assay (ELISA) for the measurement of canine S100A12

Veterinary Clinical Pathology
Romy M HeilmannJ M Steiner

Abstract

Canine S100 calcium-binding protein A12 (cS100A12) shows promise as biomarker of inflammation in dogs. A previously developed cS100A12-radioimmunoassay (RIA) requires radioactive tracers and is not sensitive enough for fecal cS100A12 concentrations in 79% of tested healthy dogs. An ELISA assay may be more sensitive than RIA and does not require radioactive tracers. The purpose of the study was to establish a sandwich ELISA for serum and fecal cS100A12, and to establish reference intervals (RI) for normal healthy canine serum and feces. Polyclonal rabbit anti-cS100A12 antibodies were generated and tested by Western blotting and immunohistochemistry. A sandwich ELISA was developed and validated, including accuracy and precision, and agreement with cS100A12-RIA. The RI, stability, and biologic variation in fecal cS100A12, and the effect of corticosteroids on serum cS100A12 were evaluated. Lower detection limits were 5 μg/L (serum) and 1 ng/g (fecal), respectively. Intra- and inter-assay coefficients of variation were ≤ 4.4% and ≤ 10.9%, respectively. Observed-to-expected ratios for linearity and spiking recovery were 98.2 ± 9.8% (mean ± SD) and 93.0 ± 6.1%, respectively. There was a significant bias between the ELISA and the RIA. ...Continue Reading

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Citations

Jun 7, 2019·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Brittany E ThamesRomy M Heilmann
Jul 28, 2020·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Angela Isabel Cabrera-GarcíaRomy M Heilmann
Jun 9, 2016·The Journal of Small Animal Practice·S M CraigJ A Lidbury
Aug 18, 2017·Reproduction in Domestic Animals = Zuchthygiene·R Hagman
Sep 18, 2018·Journal of Veterinary Internal Medicine·Romy M Heilmann, Jörg M Steiner
Jun 1, 2018·Mediators of Inflammation·Milan JovanovicIvan Jovanovic
Nov 15, 2020·The Veterinary Clinics of North America. Small Animal Practice·Juan Hernandez, Julien Rodolphe Samuel Dandrieux
Feb 9, 2021·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Anja BecherRomy M Heilmann

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