การพัฒนาปฏิกิริยาลูกโซ่โพลิเมอเรสแบบมัลติเพล็กซ์สำหรับการตรวจเชื้อ Ehrlichia canis Hepatozoon canis และ Babesia canis จากเลือดสุนัข

Chamaiporn  Nithikadpanich; Pennapa  Takam; Wasin  Charerntantanakul

Authors

Chamaiporn Nithikadpanich Program in Biotechnology, Faculty of Science, Maejo University, Chiang Mai, Thailand
Pennapa Takam Program in Biotechnology, Faculty of Science, Maejo University, Chiang Mai, Thailand
Wasin Charerntantanakul Program in Biotechnology, Faculty of Science, Maejo University, Chiang Mai, Thailand

Keywords:

Ehrlichia canis, Hepatozoon canis, Babesia canis, multiplex PCR, blood

Abstract

This study developed multiplex polymerase Chain Reaction (mPCR) from our previous study that could detect only Ehrlichia canis and Babesia canis in canine blood. In this study, we developed mPCR for simultaneous detection of VirB9 gene of E. canis and 18S rRNA gene of Hepatozoon canis and B. canis in dog blood. Infection by E. canis (rickettsia), H. canis (protozoa), and B. canis (protozoa) causes canine ehrlichiosis, hepatozoonosis, and babesiosis, respectively. These three pathogens share brown dog ticks (Rhipicephalus sanguineus) as carrier. Infected dogs usually present anemia, fever and lethargy. Current diagnostic techniques include Wright Giemsa staining of thin blood smear, PCR and immunochromatographic assay for detection of specific antibody. Among these techniques, PCR had the highest sensitivity. However, it was time-consuming, particularly when it was used for detection of individual pathogen. The mPCR technique developed in this study should save time for simultaneous detection of these three pathogens.

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