title: Detection of five avian Eimeria species by species-specific real-time polymerase chain reaction assay.
abstract: To detect five different avian Eimeria species, we applied the SYBR Green-based real-time polymerase chain reaction (PCR) assay for the diagnosis of field-isolated parasites by using their individual species-specific primer sets. The primer sets were originally designed for Eimeria acervulina, E brunetti, E necatrix, and E tenella based on the sequence of the internal transcribed spacer 1 region of ribosomal DNA, whereas for E. maxima the primer sets were derived from sequences reported previously. The detection limit of these assays was defined at 10(2) or 10(1) oocysts depending on species. Melting curves from the real-time PCR assay showed that each species has a single peak and specific melting temperature value. Fecal samples from 32 poultry farms, which were endemic for coccidiosis, were examined using this assay. The data showed that E. brunetti was found in 21 farms, E maxima and E. necatrix in 16 farms, E. tenella in 12 farms, and E. acervulina in eight farms. This survey revealed that E brunetti was highly prevalent in Japan. This technique is not only easy and rapid but also available to detect Eimeria species specifically; therefore, it can be a valuable tool for diagnostic work for chicken coccidiosis.
authors.affiliation: Nippon Institute for Biological Science, Shinmachi 9-2221-1, Ome, Tokyo 198-0024, Japan. fumiya-k@nibs.or.jp
authors.forename: Fumiya
authors.initials: F
authors.lastname: Kawahara
authors.forename: Kensuke
authors.initials: K
authors.lastname: Taira
authors.forename: Shinya
authors.initials: S
authors.lastname: Nagai
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Onaga
authors.forename: Misao
authors.initials: M
authors.lastname: Onuma
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nunoya
mesh_terms: D000818:Animals; D002645:Chickens; D003048:Coccidiosis / Q000469:parasitology / Q000662:veterinary*; D016054:DNA, Protozoan / Q000145:classification / Q000235:genetics; D004539:Eimeria / Q000145:classification* / Q000235:genetics; D005260:Female; D008297:Male; D011201:Poultry Diseases / Q000469:parasitology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D013045:Species Specificity
chemical_list: D016054:DNA, Protozoan