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A loop-mediated isothermal amplification method for rapid detection of Vibrio parahaemolyticus in seafood

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Abstract

Vibrio parahaemolyticus is an important human pathogen responsible for foodborne gastroenteritis worldwide. In this paper, a loop-mediated isothermal amplification (LAMP) method was developed for detection of V. parahaemolyticus in seafood. A set of four primers, two outer and two inner, was designed specifically to recognize the thermolabile hemolysin gene (tlh) of V. parahaemolyticus. The LAMP assay was capable of detecting a minimum of 900 fg test tube−1 for V. parahaemolyticus genomic DNA and 2.4 × 102 CFU mL−1 for pure cultures. The detection limit for the seeded seafood samples was 8.9 × 102 CFU g−1. In addition, 42 shares of natural seafood samples were tested and 8 samples were recorded positive for V. parahaemolyticus, while 6 were positive by conventional culture methods. In conclusion, the LAMP assay is an effective and low-cost method with high specificity and sensitivity for rapid detection and identification of V. parahaemolyticus both in culture isolates and seafood samples.

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Acknowledgement

This work was supported by Fok Ying Dong Education Foundation of China 114035 and Leading Academic Discipline Project of Shanghai Municipal Education Commission J50704.This work was also supported by the Maine Agricultural and Forest Experiment Station at the University of Maine with external publication number 3187.

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Authors and Affiliations

  1. College of Food Science and Technology, Shanghai Ocean University, Shanghai, 201306, People’s Republic of China

    Xiaohong Sun, Yingjie Pan, Weiqing Lan & Yong Zhao

  2. Shanghai International Airport Entry-Exit Inspection and Quarantine Bureau, Shanghai, 201202, People’s Republic of China

    Qian Xu

  3. Department of Food Science and Human Nutrition, The University of Maine, 5735 Hitchner Hall, Orono, ME, 04469–5735, USA

    Vivian C. H. Wu

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  1. Xiaohong Sun
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  2. Qian Xu
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  3. Yingjie Pan
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  4. Weiqing Lan
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  5. Yong Zhao
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  6. Vivian C. H. Wu
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Correspondence to Vivian C. H. Wu.

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Sun, X., Xu, Q., Pan, Y. et al. A loop-mediated isothermal amplification method for rapid detection of Vibrio parahaemolyticus in seafood. Ann Microbiol 62, 263–271 (2012). https://doi.org/10.1007/s13213-011-0255-0

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  • DOI: https://doi.org/10.1007/s13213-011-0255-0

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