Abstract: The phylogeny of twelve Campylobacter species was determined based on partial (1020-bp) gyrB gene sequences. Methods have been described for detection and speciation of Campylobacter, including 16S rRNA sequence analysis. However, gyrB provides a better resolution than the 16S rDNA gene for Campylobacter species with interspecies sequence similarities ranging from 58.3 to 89.2% compared to those reported for the 16S rRNA gene (ranging from 89 to 99%). A universal primer set, designed to amplify a 960-bp fragment of the gyrB gene in Campylobacter spp., was developed and used for (PCR-RFLP) of 19 strains representing twelve Campylobacter species and resulted in unique digest patterns for all twelve Campylobacter species. PCR assays for amplification of regions of the gyrB gene specific for each Campylobacter species were also developed. Using these PCR and PCR-RFLP methods results in unambiguous identification of the majority of Campylobacter species.

Abstract: The phylogeny of twelve Campylobacter species was determined based on partial (1020-bp) gyrB gene sequences. Methods have been described for detection and speciation of Campylobacter, including 16S rRNA sequence analysis. However, gyrB provides a better resolution than the 16S rDNA gene for Campylobacter species with interspecies sequence similarities ranging from 58.3 to 89.2% compared to those reported for the 16S rRNA gene (ranging from 89 to 99%). A universal primer set, designed to amplify a 900-bp fragment of the gyrB gene in Campylobacter spp., was developed and used for PCR-RFLP of 19 strains representing twelve Campylobacter species and resulted in unique digest patterns for all twelve Campylobacter species PCR assays for amplification of regions of the gyrB gene specific for each Campylobacter species were also developed. Using these PCR and PCR-RFLP methods results in unambiguous identification of the majority of Campylobacter species.

Abstract: The present invention is to provide a multiple detection method that can detect contaminating microorganisms existing in foods, including pathogenic Escherichia coli O157, Listeria monocytogenes and Salmonella spp., with high sensitivity comparable or even superior to official methods, comprising the steps of amplifying a plural number of target genes with a single PCR reaction tube and analyzing the same. The following steps are performed consecutively: (A) a step of extracting DNA of the target microorganisms to be detected by treating with at least a lytic enzyme such as Achromopepidase and Lysozyme and/or bacteriocin having lytic activity such as Enterolysine, a surfactant and a protein denaturing agent; and (B) a step of mixing a specific primer to the target microorganisms to be detected to perform multiplex PCR. Further, it is preferable to add a step of culturing with a culture condition where 1 CFU/100 g microorganisms becomes 10.sup.

Abstract: The phylogeny of twelve Campylobacter species was determined based on partial (1020-bp) gyrB gene sequences. Methods have been described for detection and speciation of Campylobacter, including 16S rRNA sequence analysis. However, gyrB provides a better resolution than the 16S rDNA gene for Campylobacter species with interspecies sequence similarities ranging from 58.3 to 89.2% compared to those reported for the 16S rRNA gene (ranging from 89 to 99%). A universal primer set, designed to amplify a 900-bp fragment of the gyrB gene in Campylobacter spp., was developed and used for PCR-RFLP of 19 strains representing twelve Campylobacter species and resulted in unique digest patterns for all twelve Campylobacter species. PCR assays for amplification of regions of the gyrB gene specific for each Campylobacter species were also developed. Using these PCR and PCR-RFLP methods results in unambiguous identification of the majority of Campylobacter species.

Abstract: Species-specific identification of campylobacters is problematic, primarily due to the absence of suitable biochemical assays for their differentiation and the existence of atypical strains. The phylogeny of twelve Campylobacter species was studied based on partial (1020-bp) gyrB gene (DNA topoisomerase beta-subunit gene) sequences. The topology of the resulting phylogenic neighbor-joining tree based on the gyrB gene was similar to that of the topology of a previously reported phylogenic tree based on the 16S rDNA gene. However, gyrB provides a better resolution than the 16S rDNA gene for Campylobacter species with interspecies sequence similarities ranging from 58.3 to 89.2%. A universal primer set, designed to amplify a 960-bp fragment of the gyrB gene in Campylobacter spp., was developed and used for PCR restriction fragment length polymorphism (PCR-RFLP) of 19 strains representing twelve Campylobacter species, including C. jejuni subsp. jejuni, C. coli, C. concisus, C. curvus, C. showae, C. mucosalis, C.

Abstract: The present invention is to provide a multiple detection method that can detect contaminating microorganisms existing in foods, including pathogenic Escherichia coli O157, Listeria monocytogenes and Salmonella spp., with high sensitivity comparable or even superior to official methods, comprising the steps of amplifying a plural number of target genes with a single PCR reaction tube and analyzing the same. The following steps are performed consecutively: (A) a step of extracting DNA of the target microorganisms to be detected by treating with at least a lytic enzyme such as Achromopepidase and Lyzocyme and/or bacteriocin having lytic activity such as Enterolysine, a surfactant and a protein denaturing agent; and (B) a step of mixing a specific primer to the target microorganisms to be detected to perform multiplex PCR.