Development of Multiplex Polymerase Chain Reaction for Detection of Ehrlichia canis, Hepatozoon canis, Babesia canis and Anaplasma platys in Dogs
Keywords:
Ehrlichia canis, Hepatozoon canis, Babesia canis, Anaplasma platys, multiplex PCRAbstract
This study developed multiplex polymerase chain reaction (mPCR) as a diagnostic technique for canine blood parasite infections. It was improved from our previous version of technique that could detect only Ehrlichia canis, Hepatozoon canis (protozoa), and Babesia canis (protozoa). In this study, we developed mPCR for simultaneous detection of VirB9 gene of E. canis, 18S rRNA gene of H. canis and B. canis, and groEL of Anaplasma platys in canine blood sample. All four pathogens require brown dog ticks (Rhipicephalus sanguineus) as carrier. Infected dogs usually present similar symptoms, i.e. anemia, anorexia, and lethargy. Current diagnostic techniques employ Wright Giemsa staining of thin blood smear, immunochromatographic assay for detection of specific antibody, and PCR. Among these techniques, PCR has the highest sensitivity, but it is time-consuming, particularly when used for detection of individual pathogen. The mPCR technique developed in this study should reduce duration of assay for simultaneous detection of these four pathogens.
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