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Background   Theileria  spp. are tick transmitted protozoa that can infect large and small ruminants causing disease and economic losses. Diagnosis of infections is often challenging, as parasites can be difficult to detect and identify microscopically and serology is unreliable. While there are PCR assays which can identify certain  Theileria  spp., there is no one PCR that has been designed to identify all recognized species that occur in ruminants and which will greatly simplify the laboratory diagnoses of infections.    Methods  Primers and probes for a genus-specific pan- Theileria  FRET-qPCR were selected by comparing sequences of recognized  Theileria  spp .  in GenBank and the test validated using reference organisms. The assay was also tested on whole blood samples from large and small ruminants from nine provinces in China.    Results  The pan- Theileria  FRET-qPCR detected all recognized species but none of the closely related protozoa. In whole blood samples from animals in China,  Theileria  spp. DNA was detected in 53.2% of the sheep tested (59/111), 44.4% of the goats (120/270) and 30.8% of the cattle (380/1,235). Water buffaloes (n = 29) were negative. Sequencing of some of the PCR products showed cattle in China were infected with  T. orientalis/T. sergenti/T. buffeli  group while  T. ovis  and  T. luwenshuni  were found in sheep and  T. luwenshuni  in goats. The prevalence of  Theileria  DNA was significantly higher in  Bos p. indicus  than in  Bos p. taurus  (77.7% vs. 18.3%) and copy numbers were also significantly higher (10 4.88  vs. 10 3.00  Theileria  18S rRNA gene copies/per ml whole blood).    Conclusions  The pan- Theileria  FRET-qPCR can detect all recognized  Theileria  spp. of ruminants in a single reaction. Large and small ruminants in China are commonly infected with a variety of  Theileria  spp.

A pan-Theileria FRET-qPCR survey for Theileria spp. in ruminants from nine provinces of China