Development and analytical validation of a radioimmunoassay for the measurement of alpha1-proteinase inhibitor concentrations in feces from healthy puppies and adult dogs.

Romy M Heilmann, Casey G Paddock, Isabelle Ruhnke, Nora Berghoff, Jan S Suchodolski, Jörg M Steiner

Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, 4474 TAMU, College Station, TX 77843-4474, USA. rheilmann@cvm.tamu.edu

Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 2011 May

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Canine α(1)-proteinase inhibitor (cα(1)-PI), a proteolysis-resistant protein with a molecular weight similar to albumin, has been shown to be clinically useful as a marker for gastrointestinal protein loss in dogs. A competitive, liquid-phase radioimmunoassay was developed and analytically validated. Fecal samples were collected from 101 healthy pet dogs of various breeds and ages, and fecal cα(1)-PI (Fcα(1)-PI) concentrations were compared between dogs of different age groups. A reference interval for Fcα(1)-PI concentration was calculated using the central 95th percentile. Analytical sensitivity of the assay was 2.2 µg Fcα(1)-PI/g feces. Observed-to-expected ratios for the serial dilution and spiking recovery of 9 and 6 fecal extracts ranged from 90.4 to 152.0% and from 71.3 to 112.3%, respectively. Coefficients of variation for intra- and interassay variability for 6 fecal extracts were ≤10.8% and ≤12.5%, respectively. The reference intervals for the mean and maximum Fcα(1)-PI from fecal samples collected on 3 consecutive days were 2.2-13.9 µg/g and 2.2-21.0 µg/g, respectively. Fcα(1)-PI was significantly higher in dogs <1 year of age (P < 0.0001 for both mean and maximum Fcα(1)-PI for the 3 samples). The radioimmunoassay described is sensitive, linear, precise, reproducible, and accurate for clinical use, thus allowing reliable quantification of Fcα(1)-PI in clinical patients. Using this assay, a mean or a maximum Fcα(1)-PI for 3 sampling days of >13.9 µg/g or >21.0 µg/g, respectively, should be considered abnormal in dogs >1 year of age. Fecal cα(1)-PI concentrations in dogs <1 year of age were significantly higher and should be carefully interpreted in this age group. © 2011 The Author(s)

Citation

Romy M Heilmann, Casey G Paddock, Isabelle Ruhnke, Nora Berghoff, Jan S Suchodolski, Jörg M Steiner. Development and analytical validation of a radioimmunoassay for the measurement of alpha1-proteinase inhibitor concentrations in feces from healthy puppies and adult dogs. Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc. 2011 May;23(3):476-85