Fig. 2

Standard curves for T. equi (a) and B. caballi (b) generated by plotting pooled Cq values from 5 separate amplifications of standards representing a 6-log dynamic range of starting template quantity (serially diluted DNA extracted from stabilates of infected equine blood) against the log of template dilution factor. Determination of the analytical sensitivity of the duplex qPCR assay for T. equi (c) and B. caballi (d). In c, standard curves were produced by amplification of serial ten-fold dilutions of a linearized plasmid containing the ema-1 gene of T. equi in the presence (gray) or absence (black) of background DNA from uninfected equine blood. Individual Cq values were plotted against the ema-1 gene copy number. In d, Cq values were plotted against the dilution factor of standards represented by DNA extracted from serial ten-fold dilutions of B. caballi-infected equine blood in uninfected equine blood (black) or DNA extracted from the undiluted sample of infected blood serially diluted in TE buffer (gray). Graphs were produced using Prism 6. Abbreviations: R², coefficient of determination; E, amplification efficiency