title: Rapid discrimination and quantification of Theileria orientalis types using ribosomal DNA internal transcribed spacers.
abstract: We report the population structure analysis of Theileria orientalis types (Ikeda, Buffeli and Chitose), the causative agent of theileriosis in cattle and its cohorts, using ITS1 and ITS2 spacers by fragment genotyping. We utilized primers flanking the two ribosomal RNA internal transcribed spacers (ITS1 and ITS2). Due to varying degrees of sequence polymorphism in the ITS regions found within and between species, we exploited the insertions and or deletions in these regions which resulted in different fragment sizes. On the basis of fragment size polymorphism, we could discriminate the three commonly found types of T. orientalis. ITS1 was capable of discriminating all three types (Ikeda-251 bp, Chitose-274 bp and Buffeli-269 bp) in one single reaction by fragment genotyping. In contrast, using ITS2, Ikeda (133-bp) a more pathogenic type was distinguishable from Buffeli/Chitose (139-bp). When compared with previous PCR detection method using, ITS1 and ITS2 genotyping was found to be more sensitive method with high specificity in population analysis and can be deployed in molecular epidemiology studies.
authors.affiliation: Department of Collaboration and Education, Research Center for Zoonosis Control, Graduate School of Veterinary Medicine, University of Hokkaido, Japan. kamau@czc.hokudai.ac.jp
authors.forename: Joseph
authors.initials: J
authors.lastname: Kamau
authors.forename: Bashir
authors.initials: B
authors.lastname: Salim
authors.forename: Naoki
authors.initials: N
authors.lastname: Yokoyama
authors.forename: Peter
authors.initials: P
authors.lastname: Kinyanjui
authors.forename: Chihiro
authors.initials: C
authors.lastname: Sugimoto
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle; D004275:DNA, Ribosomal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000032:analysis / Q000235:genetics*; D005838:Genotype; D025202:Molecular Diagnostic Techniques; D017720:Molecular Epidemiology; D058889:Molecular Typing / Q000379:methods*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D016794:Theileria / Q000145:classification* / Q000235:genetics*; D013801:Theileriasis / Q000145:classification / Q000175:diagnosis / Q000469:parasitology*
chemical_list: D004275:DNA, Ribosomal; D021903:DNA, Ribosomal Spacer