FIG. 1.
Validation of the Babesia bovis-specific nested PCR. (A) Real-time PCR detection of the B. bovis rap-1 gene (297 bp). The standard curve was generated using TOPO-RAP-1. On the x axis, the log starting quantity of template is equal to the copy number. Correlation coefficient, 0.996; slope, −3.362; intercept, 33.778; y = −3.362x + 33.778. (B) Real-time PCR detection of the B. bovis rap-1 gene from dilutions of MO7 B. bovis genomic DNA extracted from cultured infected erythrocytes. On the x axis, the log starting quantity of template is equal to genomic units. Correlation coefficient, 0.992; slope, −3.597; intercept, 33.418; y = −3.597x + 33.418. (C) Nested PCR of MO7 B. bovis genomic DNA extracted from 10-μl volumes of 50% suspensions of erythrocytes spiked with 10-fold dilutions of infected erythrocytes beginning with 108/ml. big, Babesia bigemina genomic DNA control.