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Preanalytical validation of an in‐house radioimmunoassay for measuring calprotectin in feline specimens
Author(s) -
Heilmann Romy M.,
Grützner Niels,
Handl Stefanie,
Suchodolski Jan S.,
Steiner Jörg M.
Publication year - 2018
Publication title -
veterinary clinical pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.537
H-Index - 51
eISSN - 1939-165X
pISSN - 0275-6382
DOI - 10.1111/vcp.12564
Subject(s) - radioimmunoassay , calprotectin , medicine , pathology , disease , inflammatory bowel disease
Background Calprotectin is a marker of inflammatory disorders in people, and serum and fecal calprotectin were shown to be increased in dogs with gastrointestinal inflammation. Biomarkers of gastrointestinal inflammation are currently lacking in cats. Objectives The purpose of the study was to analytically validate the canine calprotectin radioimmunoassay for quantification of calprotectin in feline specimens. Methods The immunoassay was analytically validated by determining assay working range, dilutional parallelism, spiking recovery, and intra‐ and inter‐assay variability. Reference intervals for fecal calprotectin were established from healthy cats, and the influence of age, sex, and housing condition on fecal calprotectin was determined. Results The working range of the assay was 1.5–346.2 μg/g of feces and 11.2–8654.4 μg/L of serum. Observed‐to‐expected ratios (O/E) for serial dilutions of fecal extracts ranged from 77.3% to 112.0% (mean: 99.2%) and from 95.7% to 161.4% (mean: 118.5%) for spiking recovery. Intra‐ and inter‐assay coefficients of variation for fecal samples were ≤ 11.0% and ≤ 12.8%, respectively. Fecal calprotectin concentrations ranged 1.5–66.5 μg/g (3‐day sample mean) and 1.5–126.1 μg/g (3‐day sample maximum). Housing conditions, sex, or age did not affect fecal calprotectin (all P > .05). For serial dilutions of serum samples, O/E ranged from 96.0% to 152.0% (mean: 115.7%). Serum calprotectin concentrations in healthy cats ranged from 108.8 to 255.3 μg/L (median: 158.2 μg/L). Conclusions The canine radioimmunoassay for the measurement of calprotectin is analytically sensitive, linear, reproducible, accurate, and sufficiently precise ( CV A ≤ 43.2%) for use with feline feces (with a loss of accuracy at high calprotectin concentrations). The RI s for feline fecal calprotectin are comparable to those established for dogs. Independence of fecal calprotectin from age and sex agrees with findings in dogs.