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Background  Bovine babesiosis and theileriosis is an important hemoprotozoal disease in cattles and yaks in tropical and subtropical regions leading to significant economic losses. In the field, the risk of co-infection between the bovine  Babesia  and  Theileria  species is very high. Thus, it is necessary to develop a simple, accurate, rapid and cost-effective method for large-scale epidemic investigation, in particular for the detection of co-infection in field.    Methods  In this study, DNA sequences of a ribosomal protein S8 (RPS8) gene from eight species of cattle piroplasms in China were used to develop a species-specific PCR-RFLP diagnostic tool. The eight  Theileria  and  Babesia  species could be differentiated by digesting the RPS8 PCR product with M bo  I.    Results  The sensitivity of the PCR assays was 0.1 pg DNA for  Babesia  species but 1 pg DNA for  Theileria  species. The clearly different size of the PCR-RFLP products allowed for a direct discrimination between eight bovine  Theileria  and  Babesia  species ( T. annulata ,  T. sinensis ,  T. sergenti ,  B. ovata ,  B. bovis ,  B. bigemina ,  B. major  and  Babesia  species Kashi isolate).    Conclusion  Our results indicated that the established method based on the RPS8 gene was a reliable molecular diagnostic tool for the simultaneous detection and identification of bovine  Babesia  and  Theileria  species in China, which could be applicable for the survey of parasite dynamics, epidemiological studies as well as prevention and control of the disease.

A PCR-RFLP Assay targeting RPS8 gene for the discrimination between bovine Babesia and Theileria species in China