Development and analytical validation of an enzyme-linked immunosorbent assay (ELISA) for the measurement of alpha(1)-proteinase inhibitor in serum and faeces from cats.

TitleDevelopment and analytical validation of an enzyme-linked immunosorbent assay (ELISA) for the measurement of alpha(1)-proteinase inhibitor in serum and faeces from cats.
Publication TypeJournal Article
Year of Publication2012
AuthorsBurke KF, Ruaux CG, Suchodolski JS, Williams DA, Steiner JM
JournalResearch in veterinary science
Volume93
Issue2
Pagination995-1000
Date Published2012 Oct
ISSN1532-2661
Keywordsalpha 1-Antitrypsin, Animals, Cats, Enzyme-Linked Immunosorbent Assay, Feces, Reproducibility of Results, Sensitivity and Specificity
Abstract

The objective of this study was to develop and analytically validate an ELISA for the measurement of alpha(1)-proteinase inhibitor (α(1)-PI) in serum and faeces from cats. Lower detection limit, linearity, accuracy, precision, reproducibility, and reference intervals were determined. The lower detection limits were 0.02 g/L for serum and 0.04 μg/g for faeces. The observed-to-expected (O/E) ratios for serial dilutions of serum and faecal samples ranged from 100.0 to 129.7% (mean±SD: 112.2±9.9%) and 103.5 to 141.6% (115.6±12.8%), respectively. The O/E ratios for samples spiked with seven known concentrations of α(1)-PI ranged from 82.3 to 107.8% (94.7±7.6%) for serum, and 78.5 to 148.7% (96.8±18.2%) for faeces. The coefficients of variation for intra-assay and inter-assay variability were <7.9% and <12.1% for serum, and 5.3%, 11.8%, 14.2%, and 7.7%, 10.2%, 20.4% for faeces, respectively. Reference intervals were 0.6-1.4 g/L for serum and upto 1.6 μg/g for faeces. We conclude that this ELISA is sufficiently linear, accurate, precise, and reproducible for clinical evaluation.

DOI10.1016/j.rvsc.2011.10.012
Alternate JournalRes. Vet. Sci.