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.
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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.
author list (cited authors)
Burke, K. F., Ruaux, C. G., Suchodolski, J. S., Williams, D. A., & Steiner, J. M.
complete list of authors
Burke, KF||Ruaux, CG||Suchodolski, JS||Williams, DA||Steiner, JM