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Research Article

ScienceAsia 30 (2004): 67-73 |doi: 10.2306/scienceasia1513-1874.2004.30.067

Specific and Highly Sensitive Primers for PCR Detection of Babesia bovis

Nutcha Patarapadungkit,a Suporn Nuchadomrong,a Nison Sattayasai,a,* Patchima Indrakamhangb and Sakda Daduanga


ABSTRACT: Various portions of the published sequence of Babesia bovis mitochondrial DNA were selected for detection of B. bovis by PCR-based methods. Five primer pairs, namely P1, P2, P3, P4 and P5, were designed from the gene sequence. Only P4 and P5 yielded PCR products of B. bovis DNA but not of bovine DNA. The PCR products are 120 and 181 bp for P4 and P5, respectively. The specificity of PCR was examined using DNA template from various haemoparasites: B. bovis, Trypanosoma spp., B. bigemina, Theileria spp., Anaplasma marginale and A. centrale. P5 was specific for B. bovis whereas P4 showed cross-amplification with A. centrale DNA. The P5 primer pair, which was designed from the region closed to mitochondrial large subunit ribosomal DNA of B. bovis, showed sensitivity equivalent to the Fahrimal,s primers but with higher amount of PCR product. The better result of PCR using this primer pair was confirmed by the duplex PCR when amplification of the apocytochrome b region was performed concurrently. Our study provides an alternative choice for PCR detection of B. bovis and demonstrates another region within mitochondrial DNA which is also the valuable target for specific amplification of the parasite DNA.

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a Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand.
b Immuno-Serology Section, National Institute of Animal Health, Department of Livestock Development,

* Corresponding author, E-mail: nison@kku.ac.th

Received 25 May 2003, Accepted 12 Nov 2003