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Ovine babesiosis and theileriosis are important hemoprotozoal diseases of sheep and goats in tropical and subtropical regions that lead to economic losses in these animals.PCR: restriction fragment length polymorphism (PCR:RFLP) is a reliable molecular diagnostic tool for discriminating Theileria or Babesia species in the same host.In this study,The DNA sequences of a ribosomal protein S8 (RPS8) gene from four species ofpiroplasms in China were used to develop a species:specific PCR:RFLP diagnostic tool.The sensitivity of the PCR assays was 0.1 pg DNA for B.motasi and 1 pg DNA for T.uilenbergi and 10 pg DNA for Babesia sp.Xinjiang:2005 and T.luwenshuni.The clear size difference of the PCR products allowed for a direct discrimination for B.motasi,Babesia sp.Xinjiang:2005 and ovine Theileria species (T.uilenbergi and T.luwenshuni),except that the mixed infection between T.uilenbergi and T.luwenshuni may be difficult to distinguish,simply after the electrophoretic separation of the amplification products.Further T.uilenbergi and T.luwenshuni diagnoses were made by digesting the PCR product with Sac I.The established method could be applicable for the survey of parasite dynamics,and epidemiological studies as well as prevention and control of the disease.