Abstract
Vibrio cholerae and V. vulnificus are of major concern due to their effect on public health throughout the world. It is therefore imperative to identify a gene and method that are suitable for the accurate species-specific detection of these two species. A duplex polymerase chain reaction (PCR) assay was developed using two sets of primers targeting the groEL gene for the accurate simultaneous detection of V. cholerae and V. vulnificus. The nucleotide sequence of the groEL gene was compared with the sequences of other Vibrio and non-Vibrio species. The specificity of two primer sets for duplex PCR was checked using 24 Vibrio and 8 non-Vibrio species. The primer sets were found to be specific for these two species and could detect both of the target bacterial species without any ambiguity, even when comparing closely related species. For both species, the detection limit was 100 pg of purified genomic DNA. The duplex PCR showed high specificity and sensitivity for each species and was sufficient for the detection of V. cholerae and V. vulnificus from artificially infected shellfish tissue, flounder, and even inoculated seawater. This method is simple and cost-effective, and can be utilized for the simultaneous detection of both species, thus representing an effective tool for both epidemiologist and ecologist.
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Acknowledgments
This work was supported by a grant from the National Fisheries Research and Development Institute (NFRDI), Republic of Korea. Y.-R. Kim also wishes to thank Novus International, Inc. for financial support in the form of a scholarship.
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Hossain, M.T., Kim, YR., Kim, EY. et al. Detection of Vibrio cholerae and Vibrio vulnificus by duplex PCR specific to the groEL gene. Fish Sci 79, 335–340 (2013). https://doi.org/10.1007/s12562-012-0586-1
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DOI: https://doi.org/10.1007/s12562-012-0586-1