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Isolation of the groESL cluster from Vibrio anguillarum and PCR detection targeting groEL gene

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  • Aquaculture
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Abstract

Vibrio anguillarum is a major pathogenic bacterium that causes vibriosis and septicemia in fish and shellfish. In this study, we identified the groESL genes, which encode bacterial chaperonins, from V. anguillarum. The groE gene cluster consisted of a 291-bp groES gene, a 69-bp intergenic spacer region, and a 1,635-bp groEL gene order. Sequence analysis with the groESL gene of Vibrio species exhibited that the groEL gene was more species-specific and suitable than the groES gene for V. anguillarum detection. Owing to the difficulty in distinguishing V. anguillarum from the closely related V. ordalii, we compared the sequences of groEL from V. anguillarum and the groEL homolog hsp60 from V. ordalii, in order to design a primer set based on a region dissimilar between the two. PCR with the groEL primer set produced a clear 195-bp amplicon in six serotypes of V. anguillarum, whereas 23 Vibrio species of 39 samples, including V. ordalii, and 10 species of enteric bacteria gave no bands. PCR using the groEL primers also amplified a unique product from V. anguillarum-infected flounder and oyster tissues. These results demonstrate that the groEL-target PCR assay is a sensitive and species-specific tool for the detection of V. anguillarum infection.

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Acknowledgments

This research was supported by a project grant (YSG-RE0701) from Yeongnam Sea Grant, Korea.

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Authors and Affiliations

  1. Department of Biotechnology, Pukyong National University, Busan, 608-737, Korea

    Dong-Gyun Kim, Yu-Ri Kim, Eun-Young Kim, Hyun Min Cho & In-Soo Kong

  2. Division of Infectious Diseases, Department of Medicine, Duke University Medical Center, Durham, NC, 27710, USA

    Sun-Hee Ahn

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  1. Dong-Gyun Kim
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  2. Yu-Ri Kim
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  3. Eun-Young Kim
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  4. Hyun Min Cho
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  5. Sun-Hee Ahn
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Correspondence to In-Soo Kong.

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Kim, DG., Kim, YR., Kim, EY. et al. Isolation of the groESL cluster from Vibrio anguillarum and PCR detection targeting groEL gene. Fish Sci 76, 803–810 (2010). https://doi.org/10.1007/s12562-010-0266-y

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  • DOI: https://doi.org/10.1007/s12562-010-0266-y

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