Abstract
BackgroundThe genus Vibrio is clinically significant and major pathogenic Vibrio species causing human Vibrio infections are V. cholerae, V. parahaemolyticus, V. vulnificus, V. alginolyticus and V. mimicus. In this study, we screened for novel genetic markers using comparative genomics and developed a Vibrio multiplex PCR for the reliable diagnosis of the Vibrio genus and the associated major pathogenic Vibrio species.MethodsA total of 30 Vibrio genome sequences were subjected to comparative genomics, and specific genes of the Vibrio genus and five major pathogenic Vibrio species were screened. The designed primer sets from the screened genes were evaluated by single PCR using DNAs from various Vibrio spp. and other non-Vibrio bacterial strains. A sextuplet multiplex PCR using six primer sets was developed to enable detection of the Vibrio genus and five pathogenic Vibrio species.ResultsThe designed primer sets from the screened genes yielded specific diagnostic results for target the Vibrio genus and Vibrio species. The specificity of the developed multiplex PCR was confirmed with various Vibrio and non-Vibrio strains. This Vibrio multiplex PCR was evaluated using 117 Vibrio strains isolated from the south seashore areas in Korea and Vibrio isolates were identified as Vibrio spp., V. parahaemolyticus, V. vulnificus and V. alginolyticus, demonstrating the specificity and discriminative ability of the assay towards Vibrio species.ConclusionsThis novel multiplex PCR method could provide reliable and informative identification of the Vibrio genus and major pathogenic Vibrio species in the food safety industry and in early clinical treatment, thereby protecting humans against Vibrio infection.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-015-0577-3) contains supplementary material, which is available to authorized users.
Highlights
The genus Vibrio is clinically significant and major pathogenic Vibrio species causing human Vibrio infections are V. cholerae, V. parahaemolyticus, V. vulnificus, V. alginolyticus and V. mimicus
1256 genes expected to be present in the Vibrio genus were selected based on the outputs of the Basic Local Alignment Search Tool (BLAST) program that indicated their relatively high matched DNA length, thereby eliminating genes of low homology within the Vibrio genus
These genes were compared to the non-redundant database of National Center for Biotechnology Information (NCBI) to eliminate genes that were highly matched with other biological sources
Summary
The genus Vibrio is clinically significant and major pathogenic Vibrio species causing human Vibrio infections are V. cholerae, V. parahaemolyticus, V. vulnificus, V. alginolyticus and V. mimicus. The Vibrio genus, which consists of more than 30 species, includes a number of major foodborne pathogens Eleven of these Vibrio species are known to be human pathogens causing toxigenic cholera and other infections (vibriosis). Molecular biological DNA-based diagnostic methods, especially polymerase chain reaction (PCR), have been studied and developed for accurate and rapid identification of Vibrio spp. These methods provide advantages relative to and/or that complement standard microbiological culture-based methods [8, 11]. Kim et al BMC Microbiology (2015) 15:239
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