Analysis of different RAPD primers to discriminate Babesia bovis strains

BARAVALLE, ME.; THOMPSON, C; FLORÍN CHRISTENSEN, M; TORIONI DE ECHAIDE, S.; ECHAIDE, I

Palermo (Italia)

Simposio; Second Babesia World Summit; 2007

Instituto Zooprofilattico Sperimentale della Sicilia

Molecular markers to characterize Babesia bovis isolates constitute a useful tool for epidemiological studies, to understand the dynamic of infection and to monitor the stability of vaccine strains. In this study, we have started to evaluate the use of the random amplification of polymorphic DNA (RAPD) technique for isolate characterization. Six 10 bp oligonucleotides previously used to characterize other Apicomplexan protozoan, were selected and the band patterns obtained for two different phenotype of B. bovis strains were analyzed and compared with those obtained for two other hemoparasites: B. bigemina and Anaplasma marginale and for bovine DNA.The size of the amplicons obtained with oligonucleotides ILO 509, ILO 872 and B12 are summarized in PCR amplification using ILO509 and ILO872 oligonucleotides yielded B. bovis specific bands that could clearly differentiate between BboR1A and BboS2P strains. Using B12 it was possible to differentiate B. bovis from B. bigemina, but was not possible to distinguish between strains BboR1A and BboS2P.Ongoing works are directed to assess new oligonucleotides and sequencing of cloned amplicons in order to design suitable primers to define specific molecular markers for typification of B. bovis strains.