A Novel and High Throughput Methods for Detection and Identification of Theileria equi and Babesia caballi

Two methods; namely, high-resolution melting (HRM) analysis and capillary electrophoresis were used to detect and differentiate between Theileria equi and Babesia caballi parasites that cause equine piroplasmosis worldwide. The HRM was evaluated using control samples from the United States Department of Agriculture and field samples collected from horses in Sudan and China. A region of the 18S rRNA gene, with four known nucleotide differences between the two parasites, was selected for primer design. HRM analysis successfully allowed detection and differentiation of T. equi and B. caballi without the necessity of performing time-consuming and expensive post-PCR procedures such as sequencing or restriction digestion. In another experiment, capillary electrophoresis was used to detect and differentiate between T. equi and B. caballi, using one set of primers designed to amplify the 18S rRNA gene in a single PCR. Capillary electrophoresis method was found to be powerful in detecting mixed infections in artificially mixed control samples. The results suggested that these two methods could be ideal approaches for rapid genotyping, which is required to determine a drug of choice or to administer an appropriate vaccine during an outbreak. The data obtained in this study may contribute to the development of a national control strategy of EP in the Sudan