title: Genetic analysis and development of species-specific PCR assays based on ITS-1 region of rRNA in bovine Eimeria parasites.
abstract: At present, morphological characteristics of oocyst is the only achievable method for the identification of bovine coccidia to the species level. In this study, the internal transcribed spacer 1 (ITS-1) region of ribosomal RNA genes of six bovine Eimeria species; E. alabamensis, E. auburnensis, E. bovis, E. cylindrica, E. ellipsoidalis and E. zuernii, were sequenced and analyzed the phylogenetic relationship among them. In pair-wise alignment, the sequences among the same species had high homology of over 90%. E. bovis and E. zuernii were closely related within the same cluster. This cluster and E. alabamensis were distant from major cluster of bovine coccidia that included E. auburnensis, E. cylindrica and E. ellipsoidalis. Species-specific PCR assays based on the amplification of the ITS-1 region were also developed to identify the 6 pathogens. The ITS-1 region of each Eimeria species had sufficient inter-specific sequence variation enough to design the primer sets that differentially amplified each target species. This PCR assay for the detection and differentiation of Eimeria parasite showed higher sensitivity when compared to the conventional oocyst-morphological examination. This is the first attempt for the identification of 6 bovine Eimeria parasites in the genomic level and may provide as useful methods for diagnosis and epidemiology of bovine coccidial infection.
authors.affiliation: Nippon Institute for Biological Science, Shin-machi 9-2221-1, Ome, Tokyo 198-0024, Japan. fumiya-k@nibs.or.jp
authors.forename: Fumiya
authors.initials: F
authors.lastname: Kawahara
authors.forename: Guohong
authors.initials: G
authors.lastname: Zhang
authors.forename: Claro N
authors.initials: CN
authors.lastname: Mingala
authors.forename: Yu
authors.initials: Y
authors.lastname: Tamura
authors.forename: Masateru
authors.initials: M
authors.lastname: Koiwa
authors.forename: Misao
authors.initials: M
authors.lastname: Onuma
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nunoya
mesh_terms: D000818:Animals; D002417:Cattle; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D004539:Eimeria / Q000145:classification* / Q000235:genetics*; D008969:Molecular Sequence Data; D033661:Oocysts / Q000166:cytology; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D021903:DNA, Ribosomal Spacer