title: Simultaneous identification of three highly pathogenic Eimeria species in rabbits using a multiplex PCR diagnostic assay based on ITS1-5.8S rRNA-ITS2 fragments.
abstract: Eimeria stiedai, E. intestinalis, and E. flavescens are highly pathogenic in rabbits, especially rabbits younger than 3 months. In this study, the complete ITS1-5.8S rRNA-ITS2 sequences of six rabbit Eimeria species, E. stiedai, E. intestinalis, E. flavescens, E. media, E. magna, and E. irresidua, were cloned with universal primers for the genus Eimeria and genomic DNA of LY and KF isolates as templates. These results revealed that both ITS1 and ITS2 sequences were specific to each Eimeria species in rabbits. A specific and sensitive multiplex PCR diagnostic assay based on polymorphic sites of ITS1 and ITS2 was developed and used to identify the three highly pathogenic species from rabbits, E. stiedai, E. intestinalis, and E. flavescens. Our findings provide a powerful tool for the clinical differentiation of highly pathogenic Eimeria species in rabbits and the study of the population genetics of rabbit coccidia.
authors.affiliation: Animal Quarantine Laboratory, College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China. ywchao11@126.com
authors.forename: Wenchao
authors.initials: W
authors.lastname: Yan
authors.forename: Wenlong
authors.initials: W
authors.lastname: Wang
authors.forename: Tianqi
authors.initials: T
authors.lastname: Wang
authors.forename: Xun
authors.initials: X
authors.lastname: Suo
authors.forename: Weifeng
authors.initials: W
authors.lastname: Qian
authors.forename: Shuai
authors.initials: S
authors.lastname: Wang
authors.forename: Di
authors.initials: D
authors.lastname: Fan
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D003048:Coccidiosis / Q000469:parasitology / Q000662:veterinary*; D004275:DNA, Ribosomal / Q000235:genetics*; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D004539:Eimeria / Q000145:classification / Q000472:pathogenicity*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011817:Rabbits; D013045:Species Specificity; D014774:Virulence
chemical_list: D004275:DNA, Ribosomal; D021903:DNA, Ribosomal Spacer