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Identification of Trypanosoma evansi by DNA hybridisation using a non-radioactive probe generated by arbitrary primer
S H Basagoudanavar1, J R Rao, S Omanwar
1National Biotechnology Centre, Indian Veterinary Research Institute, Izatnagar 243 122, India.
Abstract:
A highly reproducible, dominant, monomorphic fragment of 473 base pair (bp) amplified from the genome of Trypanosoma evansi by arbitrary primer-polymerase chain reaction (AP-PCR) was labelled with digoxigenin and investigated for its potential as DNA probe. Dot-blot hybridisation of total genomic DNA with the probe proved useful in detecting bubaline, cameline and equine strains of T. evansi down to 10 pg of parasite template DNA. No cross-hybridisation was seen with Babesia bigemina, Theileria annulata and the bubaline host DNA. This probe may facilitate laboratory identification of T. evansi in developing countries, without the inherent risk associated with radioisotopes.