title: Evaluation of a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for the identification of Campylobacter strains isolated from poultry in Thailand.
abstract: We have recently developed a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for identifying Campylobacter jejuni, C. coli and C. fetus. In the present study, the applicability of this assay was evaluated with 34 Campylobacter-like organisms isolated from poultry in Thailand for species identification and was compared with other assays including API Campy, 16S rRNA gene sequence, and hippuricase (hipO) gene detection. Of the 34 strains analyzed, 20, 10 and 1 were identified as C. jejuni, C. coli, and Arcobacter cryaerophilus, respectively, and 3 could not be identified by API Campy. However, 16S rRNA gene analysis, showed that all 34 strains are C. jejuni/coli. To discriminate between these 2 species, the hipO gene, which is specifically present in C. jejuni, was examined by PCR and was detected in 20 strains, which were identified as C. jejuni by API Campy but not in the remaining 14 strains. Collective results indicated that 20 strains were C. jejuni whereas the 14 strains were C. coli. When the cdt gene-based multiplex PCR was employed, however, 19, 20 and 19 strains were identified as C. jejuni while 13, 14 and 13 were identified as C. coli by the cdtA, cdtB and cdtC gene-based multiplex PCR, respectively. Pulsed-field gel electrophoresis revealed that C. jejuni and C. coli strains analyzed are genetically diverse. Taken together, these data suggest that the cdt gene-based multiplex PCR, particularly cdtB gene-based multiplex PCR, is a simple, rapid and reliable method for identifying the species of Campylobacter strains.
authors: Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai, Osaka, Japan.
authors: Worada
authors: W
authors: Samosornsuk
authors: Masahiro
authors: M
authors: Asakura
authors: Emi
authors: E
authors: Yoshida
authors: Takashi
authors: T
authors: Taguchi
authors: Kazuhiko
authors: K
authors: Nishimura
authors: Boonchuay
authors: B
authors: Eampokalap
authors: Vongsavanh
authors: V
authors: Phongsisay
authors: Wanpen
authors: W
authors: Chaicumpa
authors: Shinji
authors: S
authors: Yamasaki
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015373:Bacterial Typing Techniques / Q000379:methods*; D002167:Campylobacter / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D002169:Campylobacter Infections / Q000382:microbiology / Q000662:veterinary*; D017931:DNA Primers; D016521:Electrophoresis, Gel, Pulsed-Field; D005798:Genes, Bacterial; D014644:Genetic Variation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011200:Poultry; D011201:Poultry Diseases / Q000382:microbiology*; D013785:Thailand
chemical_list: D001427:Bacterial Toxins; D017931:DNA Primers; C067044:cytolethal distending toxin