title: Real-time polymerase chain reaction (PCR) assays for the specific detection and quantification of seven Eimeria species that cause coccidiosis in chickens.
abstract: Coccidiosis of chickens is an economically important disease caused by infection with species of Eimeria. The oocysts of some of the seven recognized species are difficult to distinguish morphologically and for this reason diagnostic laboratories are increasingly utilizing DNA-based technologies for the specific identification of Eimeria. The real-time PCR provides both sensitivity and speed for the analysis of DNA samples, and the approach has the capability of quantifying DNA. Together with a protocol for the extraction of DNA directly from faecal samples, real-time PCR assays have been established for the detection and quantification of seven species of Eimeria that infect chickens in Australia. The assays target one genetic marker, the second internal transcribed spacer of nuclear ribosomal DNA (ITS-2), use TaqMan MGB technology with species-specific probes, and can be multiplexed in pairs such that the seven species of Eimeria can be screened in four reaction tubes. A test screen of commercial flocks identified more Eimeria-infected chickens than were detected by coproscopic examination for oocysts. These molecular assays can also be used for the quality control of mixed-species vaccines. The ability to multiplex the assays makes them particularly practical for screening samples from chickens with mixed-species infections where the relative abundance of each Eimeria species present is required.
authors.affiliation: Department of Primary Industries & Fisheries, Animal Research Institute, LMB No 4, Moorooka, Queensland 4105, Australia. jessica.morgan@dpi.qld.gov.au
authors.forename: J A T
authors.initials: JA
authors.lastname: Morgan
authors.forename: G M
authors.initials: GM
authors.lastname: Morris
authors.forename: B M
authors.initials: BM
authors.lastname: Wlodek
authors.forename: R
authors.initials: R
authors.lastname: Byrnes
authors.forename: M
authors.initials: M
authors.lastname: Jenner
authors.forename: C C
authors.initials: CC
authors.lastname: Constantinoiu
authors.forename: G R
authors.initials: GR
authors.lastname: Anderson
authors.forename: A E
authors.initials: AE
authors.lastname: Lew-Tabor
authors.forename: J B
authors.initials: JB
authors.lastname: Molloy
authors.forename: R B
authors.initials: RB
authors.lastname: Gasser
authors.forename: W K
authors.initials: WK
authors.lastname: Jorgensen
mesh_terms: D000818:Animals; D002645:Chickens; D003048:Coccidiosis / Q000469:parasitology / Q000662:veterinary*; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D004539:Eimeria / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000469:parasitology*; D015203:Reproducibility of Results
chemical_list: D021903:DNA, Ribosomal Spacer