title: Development and analytical validation of a radioimmunoassay for the measurement of alpha1-proteinase inhibitor concentrations in feces from healthy puppies and adult dogs.
abstract: 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.
authors: Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, 4474 TAMU, College Station, TX 77843-4474, USA. rheilmann@cvm.tamu.edu
authors: Romy M
authors: RM
authors: Heilmann
authors: Casey G
authors: CG
authors: Paddock
authors: Isabelle
authors: I
authors: Ruhnke
authors: Nora
authors: N
authors: Berghoff
authors: Jan S
authors: JS
authors: Suchodolski
authors: Jörg M
authors: JM
authors: Steiner
mesh_terms: D000367:Age Factors; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D004283:Dog Diseases / Q000175:diagnosis; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005243:Feces / Q000737:chemistry*; D005767:Gastrointestinal Diseases / Q000175:diagnosis / Q000662:veterinary; D011863:Radioimmunoassay / Q000379:methods / Q000662:veterinary*; D012016:Reference Values; D015203:Reproducibility of Results; D000515:alpha 1-Antitrypsin / Q000032:analysis*
chemical_list: D015415:Biomarkers; D000515:alpha 1-Antitrypsin