title: A PCR-RFLP assay for the detection and differentiation of Campylobacter jejuni, C. coli, C. fetus, C. hyointestinalis, C. lari, C. helveticus and C. upsaliensis.
abstract: Although Campylobacter jejuni and Campylobacter coli are the most common bacterial causes of human gastrointestinal diseases, other Campylobacter species are also involved in human and animal infections. In this study, we developed a cytolethal distending toxin (cdt) gene-based PCR-RFLP assay for the detection and differentiation of C. jejuni, C. coli, C. fetus, C. hyointestinalis, C. lari, C. helveticus and C. upsaliensis. Previously designed common primers, which can amplify the cdtB gene of C. jejuni, C. coli and C. fetus, were used for detecting seven Campylobacter species and differentiating between them by restriction digestion. The PCR-RFLP assay was validated with 277 strains, including 35 C. jejuni, 19 C. coli, 20 C. fetus, 24 C. hyointestinalis, 13 C. lari, 2 C. helveticus, 22 C. upsaliensis, 3 other Campylobacter spp. and 17 other species associated with human diseases. Sensitivity and specificity of the PCR-RFLP assay were 100 % except for C. hyointestinalis (88 % sensitivity). Furthermore, the PCR-RFLP assay successfully detected and differentiated C. jejuni, C. coli and C. fetus in clinical and animal samples. The results indicate that the PCR-RFLP assay is useful for the detection and differentiation of seven Campylobacter species important for human and animal diseases.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Osaka University, Japan.|Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Japan.
authors.forename: Kazumasa
authors.initials: K
authors.lastname: Kamei
authors.affiliation: Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Asakura
authors.affiliation: Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Japan.
authors.forename: Srinuan
authors.initials: S
authors.lastname: Somroop
authors.affiliation: Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Japan.
authors.forename: Noritoshi
authors.initials: N
authors.lastname: Hatanaka
authors.affiliation: Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Hinenoya
authors.affiliation: Department of Pediatrics, Mizushima General Hospital, Okayama, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Nagita
authors.affiliation: Faculty of Agriculture, University of Miyazaki, Japan.
authors.forename: Naoaki
authors.initials: N
authors.lastname: Misawa
authors.affiliation: Graduate School of Environmental Health Sciences, Azabu University, Kanagawa, Japan.
authors.forename: Motoo
authors.initials: M
authors.lastname: Matsuda
authors.affiliation: Graduate School of Pharmaceutical Sciences, Osaka University, Japan.
authors.forename: Shinsaku
authors.initials: S
authors.lastname: Nakagawa
authors.affiliation: Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Yamasaki
mesh_terms: D000293:Adolescent; D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics; D002167:Campylobacter / Q000145:classification* / Q000302:isolation & purification*; D002169:Campylobacter Infections / Q000175:diagnosis* / Q000382:microbiology*; D002417:Cattle; D002648:Child; D002675:Child, Preschool; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D001427:Bacterial Toxins; D017931:DNA Primers; D004269:DNA, Bacterial; C067044:cytolethal distending toxin