Rapid identification of thermotolerant Campylobacter jejuni, Campylobacter coli, Campylobacter lari, and Campylobacter upsaliensis from various geographic locations by a GTPase-based PCR-reverse hybridization assay.
Recently, a gene from Campylobacter jejuni encoding a putative GTPase was identified. Based on two semiconserved GTP-binding sites encoded within this gene, PCR primers were selected that allow amplification of a 153-bp fragment from C. jejuni, C. coli, C. lari, and C. upsaliensis. Sequence analysis of these PCR products revealed consistent interspecies variation, which allowed the definition of species-specific probes for each of the four thermotolerant Campylobacter species. Multiple probes were used to develop a line probe assay (LiPA) that permits analysis of PCR products by a single reverse hybridization step. A total of 320 reference strains and clinical isolates from various geographic origins were tested by the GTP-based PCR-LiPA. The PCR-LiPA is highly specific in comparison with conventional identification methods, including biochemical and whole-cell protein analyses. In conclusion, a simple method has been developed for rapid and highly specific identification of thermotolerant Campylobacter species.
D001315:Australia; D001483:Base Sequence; D002167:Campylobacter / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D002169:Campylobacter Infections / Q000382:microbiology*; D017000:Campylobacter coli / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D016123:Campylobacter jejuni / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D020558:GTP Phosphohydrolases / Q000235:genetics*; D005798:Genes, Bacterial; D014644:Genetic Variation; D005843:Geography; D006358:Hot Temperature; D006801:Humans; D008969:Molecular Sequence Data; D009426:Netherlands; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013557:Switzerland; D014481:United States