Development and analytical validation of an enzyme-linked immunosorbent assay (ELISA) for the measurement of alpha1-proteinase inhibitor in serum and faeces from cats

K. F. Burke, C. G. Ruaux, J. S. Suchodolski, David A Williams, J. M. Steiner

Research output: Contribution to journalArticlepeer-review

Abstract

The objective of this study was to develop and analytically validate an ELISA for the measurement of alpha1-proteinase inhibitor (α1-PI) in serum and faeces from cats. Lower detection limit, linearity, accuracy, precision, reproducibility, and reference intervals were determined. The lower detection limits were 0.02g/L for serum and 0.04μg/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 α1-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 0.6-1.4g/L for serum and upto 1.6μg/g for faeces. We conclude that this ELISA is sufficiently linear, accurate, precise, and reproducible for clinical evaluation.

Original languageEnglish (US)
Pages (from-to)995-1000
Number of pages6
JournalResearch in Veterinary Science
Volume93
Issue number2
DOIs
StatePublished - Oct 1 2012

Keywords

  • Alpha-PI
  • Cats
  • ELISA
  • Faeces
  • Serum
  • Validation

ASJC Scopus subject areas

  • veterinary(all)

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