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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abstract
The objective of this study was to develop and analytically validate an ELISA for the measurement of alpha(1)-proteinase inhibitor ((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 g/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% (meanSD: 112.29.9%) and 103.5 to 141.6% (115.612.8%), respectively. The O/E ratios for samples spiked with seven known concentrations of (1)-PI ranged from 82.3 to 107.8% (94.77.6%) for serum, and 78.5 to 148.7% (96.818.2%) 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 g/g for faeces. We conclude that this ELISA is sufficiently linear, accurate, precise, and reproducible for clinical evaluation.
