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Background  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 . 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.    Methods  A 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.    Results  The 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.    Conclusions  This 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 material  The online version of this article (doi:10.1186/s12866-015-0577-3) contains supplementary material, which is available to authorized users.

Multiplex PCR for detection of the Vibrio genus and five pathogenic Vibrio species with primer sets designed using comparative genomics