Development of multiplex polymerase chain reaction for detection of Ehrlichia canis, Babesia spp. and Hepatozoon canis in canine blood.

A multiplex polymerase chain reaction (PCR) has been developed for simul- taneous detection of canine blood parasites, Ehrlichia canis, Babesia spp and Hepatozoon canis, from blood samples in a single reaction. The multiplex PCR primers were spe- cific to E. canis VirB9, Babesia spp 16S rRNA and H. canis 16S rRNA genes. Specificity of the amplicons was confirmed by DNA sequencing. The assay was evaluated using normal canine and infected blood samples, which were detected by microscopic ex- amination. This multiplex PCR offers scope for simultaneous detection of three im- portant canine blood parasites and should be valuable in monitoring parasite infec- tions in dogs and ticks.