Rapid detection and identification of Theileria equi and Babesia caballi by high-resolution melting (HRM) analysis
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The application of high-resolution melting (HRM) analysis in the differentiation between Theileria equi and Babesia caballi 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 the 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. Our results suggest that HRM could be an ideal method for rapid genotyping, which is required to determine a drug of choice or to administer an appropriate vaccine during an outbreak.
KeywordsBabesia Melting Profile Reverse Line Blotting Babesia Species Difference Graph
This work was supported by Grant-in-Aid for JSPS fellows and for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT); the program of the Funding Research Center for Emerging and Re-emerging Infectious Disease, MEXT; and Asia-Africa S & T Strategic Cooperation Promotion Program by the Special Coordination Funds for Promoting Science & Technology, MEXT. We would like to thank Mr. Ryo Nakao for his assistance.
Conflict of interest
The authors declare that they have no competing interests.
- Dobrowolski SF, Ellingson C, Coyne T et al (2007) Mutations in the phenylalanine hydroxylase gene identified in 95 patients with phenylketonuria using novel systems of mutation scanning and specific genotyping based upon thermal melt profiles. Mol Genet Metab 91:218–227. doi: 10.1016/j.ymgme.2007.03.010 PubMedCrossRefGoogle Scholar
- Friedhoff KT, Tenter AM, Müller I (1990) Haemoparasites of equines: impact on international trade of horses. Rev Sci Tech Int Off Epizoot 9:1187–1194Google Scholar
- Kuttler KL (1988) World-wide impact of babesiosis. In: Ristic M (ed) Babesiosis of domestic animal and man. CRC, Boca Raton, pp 1–22Google Scholar
- Rhalem A, Sahibi H, Lasri S et al (2001) Validation of a competitive enzyme-linked immunosorbent assay for diagnosing Babesia equi infections of Moroccan origin and its use in determining the seroprevalence of B. equi in Morocco. J Vet Diagn Invest 13:249–251. doi: 10.1177/104063870101300311 PubMedCrossRefGoogle Scholar
- Schein E (1988) Equine babesiosis. In: Ristic M (ed) Babesiosis of domestic animal and man. CRC, Boca Raton, pp 197–208Google Scholar