Rapid identification of thermotolerant Campylobacter jejuni, Campylobactercoli, 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 i dentified. Based on two semiconserved GTP-binding sites encoded within this gene, PCR primers were selected that allow amplification of a 153-bp fragm ent from C. jejuni, C. coli, C. lari, and C. upsaliensis. Sequence analysis of these PCR products revealed consistent interspecies variation, which al lowed the definition of species-specific probes for each of the four thermo tolerant Campylobacter species. Multiple probes were used to develop a line probe assay (LiPA) that permits analysis of PCR products by a single rever se hybridization step. A total of 320 reference strains and clinical isolat es from various geographic origins were tested by the GTP-based PCR-LiPA. T he PCR-LiPA is highly specific in comparison with conventional identificati on methods, including biochemical and whole-cell protein analyses. In concl usion, a simple method has been developed for rapid and highly specific ide ntification of thermotolerant Campylobacter species.