title: A pan-Theileria FRET-qPCR survey for Theileria spp. in ruminants from nine provinces of China.
abstract: 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.
authors.forename: Yi
authors.initials: Y
authors.lastname: Yang
authors.forename: Yongjiang
authors.initials: Y
authors.lastname: Mao
authors.forename: Patrick
authors.initials: P
authors.lastname: Kelly
authors.forename: Zhangpin
authors.initials: Z
authors.lastname: Yang
authors.forename: Lu
authors.initials: L
authors.lastname: Luan
authors.forename: Jilei
authors.initials: J
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Heba S
authors.initials: HS
authors.lastname: El-Mahallawy
authors.affiliation: Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University College of Veterinary Medicine, Yangzhou 225009, Jiangsu, China. wangcm@yzu.edu.cn.
authors.forename: Chengming
authors.initials: C
authors.lastname: Wang
mesh_terms: D000818:Animals; D016054:DNA, Protozoan / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D015995:Prevalence; D012418:Ruminants / Q000469:parasitology*; D012680:Sensitivity and Specificity; D016794:Theileria / Q000145:classification / Q000302:isolation & purification*; D013801:Theileriasis / Q000097:blood / Q000453:epidemiology / Q000469:parasitology*
chemical_list: D016054:DNA, Protozoan