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

BMC Microbiology

Research Article

Ji-Oh Ryu¹  Shin-Young Lee¹  Hyun-Joong Kim¹  Ei-Seul Kim¹  Hae-Yeong Kim¹ 

[1] Institute of Life Sciences & Resources and Graduate School of Biotechnology, Kyung Hee University, 446-701, Yongin, Republic of Korea;

关键词: Identification;    PCR;    Vibrios;    V. cholerae;    V. parahaemolyticus;    V. vulnificus;    V. alginolyticus;    V. mimicus;    Comparative genomics;   

DOI  :  10.1186/s12866-015-0577-3

 received in 2015-01-08, accepted in 2015-10-19,  发布年份 2015

来源: Springer

【 摘 要 】

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.

【 授权许可】

CC BY   
© Kim et al. 2015