Veterinary Archives, Vol. 81 No. 1, 2011.
Original scientific paper
Identification of Trypanosoma vivax subtypes isolated from cattle and goats using microsatellite markers.
Savino Biryomumaisho
; Department of Veterinary Medicine, Makerere University, Kampala, Uganda
Eli Katunguka-Rwakishaya
; Department of Veterinary Medicine, Makerere University, Kampala, Uganda
George W. Lubega
; Department of Veterinary Parasitology and Microbiology, Makerere University, Kampala, Uganda
Sara E. Melville
; Department of Pathology, University of Cambridge, Cambridge, CB2 1QP, United Kingdom
Abstract
Microsatellite DNA polymorphisms can be utilised to assess intra-specific genetic diversity and hence are useful for characterisation of species and strains of trypanosomes. Here, we present four new microsatellite markers specific for T. vivax isolated from Ugandan cattle and goats. The GeneDB partial shotgun 5x coverage sequence of T. vivax available as of 1st August 2005 was used and targeted the genomic sequence of T. vivax that has no cross amplification with other livestock-infective trypanosomes. Only di-; tri-; tetra;- and pentanucleotide microsatellites not less than five units were selected. Although pentanucleotide repeats on screening appeared to have the desired variability, they gave poorer PCR products compared to di-, tri- and tetranucleotide repeats. Mononucleotide repeats presented difficulty in detecting visible bands on agarose gels from their amplification and were omitted from this study. Clear length polymorphism was obtained with guanine, thymine and adenine repeated 16 times (GTA)16 while cytosine, adenine, cytosine and thymine (CACT)15 gave size and length variability. Bands of similar size were obtained from thymine and adenine (TTA)24 microsatellite, approximately 150 base pairs long and 180-200 base pairs from the cytosine and adenine (CA)26 microsatellite. These findings suggest that different subtypes of T. vivax exist in Uganda; the polymorphic forms derived from microsatellite band size differences may suggest this parasite exhibits virulence differences as has been shown in T. Congolense subtypes.
Keywords
microsatellites; Trypanosoma vivax; subtypes
Hrčak ID:
66434
URI
Publication date:
21.2.2011.
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