The card agglutination test for dog erythrocyte antigen 1 (DEA 1) blood typing in donor dogs: Determining an appropriate cutoff to detect positivity using a receiver operating characteristic curve

Background The appropriate cutoff to define a positive point-of-care card agglutination (CA) test for dog erythrocyte antigen 1 (DEA 1) blood typing depends on whether the test can be used in the donor or recipient. Objectives By screening for CA test positivity, we aimed to evaluate the best cutoff value for DEA 1 blood typing in canine blood donors using a receiver operator characteristic (ROC) curve. Methods Ethylenediaminetetraacetic acid (EDTA) blood samples from 100 canine blood donors were blood-typed in parallel for DEA 1 using both immunochromatographic (IC) and CA tests. The effect of temperature, storage time, and anticoagulant solutions for both methods was evaluated. Unweighted and weighted Cohen's Kappa (K) statistic was calculated to evaluate the agreement between the two testing methods. The overall performance of the CA test was evaluated by generating a ROC curve using the IC test as the reference method. Results Concordant results were obtained for 86% of the samples. Unweighted and weighted K statistics demonstrated good and moderate agreement, respectively. For the CA test, the ROC curve showed an area under the curve (AUC) of 0.910, with the highest sensitivity cutoff values at >= 1+ agglutination. CA- and IC-typed EDTA blood samples stored at room temperature for up to 1 week and refrigerated for up to 1 month were concordant as were the citrate phosphate dextrose adenine 1 (CPDA-1) anticoagulated blood samples stored for up to 1 week at 4 +/- 2 degrees C. Conclusions The overall reliability of the CA method seemed to be lower than that of the IC method. When CA is used as a screening test for canine blood donors, the correct cut off is >= 1+ agglutination is recommended to maximize sensitivity