Detection of Babesia bovis in cattle by PCR-ELISA.
We established a new highly sensitive method, PCR-ELISA, for the dectection of Babesia bovis in cattle for farms in Thailand. The detection of around 2.4 x 10(-8)% parasitemia (equivalent to 1 infected erythrocyte per 2 ml) was achieved by PCR amplification followed by the ELISA detection of a biotin tagged gene. When comparing the sensitivity of PCR-ELISA with the microscopic method, our PCR-ELISA method is more sensitive than thin blood smears by at least 1,000 times. The established PCR-ELISA also showed high specificity to B. bovis with no cross reaction to other endemic parasites except for A. marginale. Regarding the detection threshold for B. bovis, the PCR-ELISA method could detect parasites inoculated into splenectomized calves at least 1 week earlier than the thin blood microscopic method. The PCR-ELISA method is a valuable screening technique for B. bovis and applicable for the routine detection of carrier states and automated analysis.
Thammasirirak
Sompong
S
Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand. somkly@kku.ac.th
Siriteptawee
Jaruwan
J
Sattayasai
Nison
N
Indrakamhang
Patchima
P
Araki
Tomohiro
T
D000818:Animals; D016793:Babesia bovis / Q000302:isolation & purification*; D001404:Babesiosis / Q000097:blood / Q000175:diagnosis / Q000662:veterinary*; D002353:Carrier State / Q000175:diagnosis / Q000662:veterinary; D002417:Cattle; D002418:Cattle Diseases / Q000097:blood / Q000175:diagnosis*; D004587:Electrophoresis, Agar Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D008853:Microscopy; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity; D013045:Species Specificity