Detection of Babesia Subspecies from Naturally Infected Dog Blood by RFLP-PCR
Blood from one year old female Boxer dog showing clinical and haematological sings of babesiosis, including fever(40°C), anaemia (RBC: 3.04/L), low haematocrit ( 21.3 percent), haemoglobinemia (7.3 g/L), was taken in Istanbul in September 2002. Examination of Giemsa-stained blood smear revealed the presence of Babesia parasites, which have morphology (pear shaped parasites) and size (4.5-5.0µm) typical of the large Babesia.
The molecular typing of the canine blood sample was performed with the PCR-RFLP assay and confirmed the presence of a B. canis vogeli subspecies in the infected dog.
Polymerase chain reaction (PCR) was first applied to diagnosis of canine babesiosis in Turkey
The 400 bp size of the piroA/piroB amplified fragment correspond to the expected size for B. canis species (whatever the subspecies involved) and as expected for the identification of B. canis vogeli, the piroA/piroB PCR fragment from Turkish canine blood sample was digested by the restriction enzyme TaqI but not Hinf I. As controls, the piroA/piroB PCR fragment B. canis rossi (South African subspecies) is digested by Hinf I but not TaqI and the piroA/piroB PCR fragment B. canis (European subspecies) is not digested by these two restriction enzymes. The PCR-RFLP assay described here to type the blood sample from Turkey revealed the presence of a B. canis vogeli subspecies in the infected dog.