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This study aimed to evaluate a commercially available real-time PCR TaqMan kit for the detection of Anaplasma marginale and Babesia bigemina. Performance of the assay was evaluated on both DNA-based plasmid and in-vitro-transcribed RNA models. Sensitivity of the assay was between 2.4 × 10(-3) (95% confidence interval: 2.3 × 10(-3) -2.7 × 10(-3) ) and 1.1 × 10(-3) (1.0 × 10(-3) -1.2 × 10(-3) ) of the expected DNA copies (B. bigemina) and RNA copies (A. marginale) per reaction, respectively. Comparisons of the assay results with those obtained by a FRET-based real-time PCR (Illumina TaqPathBabesiaFRET kit) and a parallel immunochromatographic test (Abraxis) showed good agreement (Kappa