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Use of a Granulocyte Immunofluorescence Assay Designed for Humans for Detection of Antineutrophil Cytoplasmic Antibodies in Dogs with Chronic Enteropathies

Florey, J., Viall, A., Streu, S., DiMuro, V., Riddle, A., Kirk, J., Perazzotti, L., Affeldt, K., Wagner, R., Vaden, S., Harris, T., Allenspach, K.
Journal of veterinary internal medicine 2017 v.31 no.4 pp. 1062-1066
antibodies, blood serum, confidence interval, dogs, fluorescent antibody technique, humans, kidney diseases, myeloperoxidase, seroprevalence
BACKGROUND: Perinuclear antineutrophil cytoplasmic antibodies (pANCA) previously have been shown to be serum markers in dogs with chronic enteropathies, with dogs that have food‐responsive disease (FRD) having higher frequencies of seropositivity than dogs with steroid‐responsive disease (SRD). The indirect immunofluorescence (IIF) assay used in previous publications is time‐consuming to perform, with low interobserver agreement. HYPOTHESIS/OBJECTIVES: We hypothesized that a commercially available granulocyte IIF assay designed for humans could be used to detect perinuclear antineutrophil cytoplasmic antibodies in dogs. ANIMALS: Forty‐four dogs with FRD, 20 dogs with SRD, 20 control dogs, and 38 soft‐coated wheaten terrier (SCWT) or SCWT‐cross dogs. METHODS: A granulocyte assay designed for humans was used to detect pANCA, cANCA, and antinuclear antibodies (ANA), as well as antibodies against proteinase‐3 protein (PR‐3) and myeloperoxidase protein (MPO) in archived serum samples. RESULTS: Sensitivity of the granulocyte assay to predict FRD in dogs was 0.61 (95% confidence interval (CI), 0.45, 0.75), and specificity was 1.00 (95% CI, 0.91, 1.00). A significant association was identified between positive pANCA or cANCA result and diagnosis of FRD (P < 0.0001). Agreement between the two assays to detect ANCA in the same serum samples from SCWT with protein‐losing enteropathy/protein‐losing nephropathy (PLE/PLN) was substantial (kappa, 0.77; 95% CI, 0.53, 1.00). Eight ANCA‐positive cases were positive for MPO or PR‐3 antibodies. CONCLUSIONS AND CLINICAL IMPORTANCE: The granulocyte immunofluorescence assay used in our pilot study was easy and quick to perform. Agreement with the previously published method was good.