Development of canine C-reactive protein assays

Abstract

AbstractC-reactive protein (CRP), which is released during tissue damage and inflammation, is a useful nonspecific inflammatory marker in both human and veterinary clinical practice. Veterinarians have often used human CRP assays to analyze samples from canine patients, but cross-reactivities between the species affect assay sensitivity and reliability, leading to inaccurate inflammation assessment. To improve the efficiency of inflammation assessment, we developed a canine CRP detection enzyme-linked immunosorbent assay (ELISA) for quantitative analysis and an immunochromatography assay (ICA) for semiquantitative point-of-care (POC) analysis. The ELISA demonstrated an assay detection limit of 0.5 ng/mL, quantitative linear assay range of 1.6–100 ng/mL, and intra- and inter-assay coefficient of variations of 0.7 to 10.0% and 6.0 to 9.0%, respectively; the recovery rates of samples spiked with purified canine CRP were 105 to 109%, and the parallelism assessments were 82.7 to 104.4%. The correlation between the CRP level results obtained with the ELISA and those of a currently available quantitative POC assay was 0.907 with the regression formula of y = 0.55x + 0.05. In addition, the ICA requires only 5 μL samples and a 10-min assay time, and clearly distinguished positive, weak positive, and negative samples (P < 0.001) at an approximately 5–10 µg/mL cut-off value. The developed canine CRP ELISA and ICA showed reliable assay results and a high correlation with a commercially available POC assay in clinical use. The ICA can be a useful canine CRP screening test for diagnostic purposes in veterinary clinics.