Multiplex PCR for detection of the Vibrio genus and five pathogenic Vibrio species with primer sets designed using comparative genomics. Issue 1 (December 2015)

• Record Type: Journal Article
• Authors: Kim, Hyun-Joong; Ryu, Ji-Oh; Lee, Shin-Young; Kim, Ei-Seul; Kim, Hae-Yeong
• Is Part Of: BMC microbiology. Volume 15:Issue 1(2015)
• Page Start: 1
• Page End: 11
• Publication Date: 2015-12
• Subjects: Identification -- PCR -- Vibrios -- V. cholerae -- V. parahaemolyticus -- V. vulnificus -- V. alginolyticus -- V. mimicus -- Comparative genomics; Microbiology -- Periodicals; 579.05
• DOI: 10.1186/s12866-015-0577-3
• Languages: English
• ISSNs: 1471-2180

Abstract

Abstract Background The genusVibrio is clinically significant and major pathogenicVibrio species causing humanVibrio infections areV. cholerae, V. parahaemolyticus, V. vulnificus, V. alginolyticus andV. mimicus . In this study, we screened for novel genetic markers using comparative genomics and developed aVibrio multiplex PCR for the reliable diagnosis of theVibrio genus and the associated major pathogenicVibrio species. Methods A total of 30Vibrio genome sequences were subjected to comparative genomics, and specific genes of theVibrio genus and five major pathogenicVibrio species were screened. The designed primer sets from the screened genes were evaluated by single PCR using DNAs from variousVibrio spp. and other non-Vibrio bacterial strains. A sextuplet multiplex PCR using six primer sets was developed to enable detection of theVibrio genus and five pathogenicVibrio species. Results The designed primer sets from the screened genes yielded specific diagnostic results for target theVibrio genus andVibrio species. The specificity of the developed multiplex PCR was confirmed with variousVibrio and non-Vibrio strains. ThisVibrio multiplex PCR was evaluated using 117Vibrio strains isolated from the south seashore areas in Korea andVibrio isolates were identified asVibrio spp., V. parahaemolyticus, V. vulnificus andV. alginolyticus, demonstrating the specificity and discriminative ability of the assay towardsVibrio species. Conclusions This novel multiplex PCR method could provide reliable and informative identification of theVibrio genus and major pathogenicVibrio species in the food safety industry and in early clinical treatment, thereby protecting humans againstVibrio infection.