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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
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