Theileria spp. Infect wild and domestic animals in the tropical and subtropical regions of the world. While there are many foci of ovine theileriosis in Iran, the epidemiological aspects of ovine theileriosis and theileria species that are involved in the disease are poorly understood. In this study, the use of a nested PCR for the amplification of virtually all the species of theileria is described. Fir differentiation of various theileria spp. We developed a RFLP asay as a diagnostic tool enabling direct, concurrent, highly specific and sensitive identification of theileria spp. Blood samples were collected in EDTA from 50 sheep in zabol, an area known as endemic for ovine theileriosis. Blood was used to extract DNA for PCR analysis. A 1416-bp DNA fragment was specifically amplified from blood samples of naturally infected sheep with theileria spp. For differentiation of different theileria species (T. annulata, T.lestoquardi, T. ovis) RFLP of PCR products of theileria spp. Was done. 71 percent of samples were positive for theileria spp. using nestred PCR. Analysis by RFLP showed that 14% were positive for T. ovis and 58% were positive for T. lestoquardi. This is the first report of molecular identification of theileria ovis in Iran.

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