Development of Multiplex Polymerase Chain Reaction for the Detection of Babesia spp. and Ehrlichia canis in Dog Blood

Abstract

This study developed multiplex polymerase chain reaction (multiplex PCR) for simultaneous detection of 18S rRNA gene of Babesia spp. and VirB9 gene of Ehrlichia canis in dog blood. Babesia spp. (protozoa) and E. canis (rickettsia) are the causative agents of canine babesiosis and canine Ehrlichiosis, respectively. Both pathogens have brown dog ticks (Rhipicephalus sanguineus) as carrier. Once infected, dogs manifest anemia, fever, and lethargy. Currently available diagnostic techniques include immunochromatographic test for detection of specific antibody, Wright Giemsa staining of thin blood smear, and PCR. Among these techniques, PCR has the highest sensitivity. However, it is relatively time-consuming, compared to the other two techniques, particularly when it is used for detection of individual pathogen. The multiplex PCR technique developed in this study should reduce time for the detection of both pathogens.