Development and Evaluation of a Loop-Mediated Isothermal Amplification Assay Combined with Enrichment Culture for Rapid Detection of Very Low Numbers of Vibrio parahaemolyticus in Seafood Samples

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  • Di Huiling
    College of Light Industry and Food Sciences, South China University of Technology
  • Ye Lei
    College of Light Industry and Food Sciences, South China University of Technology
  • Neogi Sucharit Basu
    College of Light Industry and Food Sciences, South China University of Technology
  • Meng Hecheng
    Graduate School of Life and Environmental Sciences, Osaka Prefecture University
  • Yan He
    College of Light Industry and Food Sciences, South China University of Technology
  • Yamasaki Shinji
    College of Light Industry and Food Sciences, South China University of Technology Graduate School of Life and Environmental Sciences, Osaka Prefecture University
  • Shi Lei
    College of Light Industry and Food Sciences, South China University of Technology

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  • Development and Evaluation of a Loop-Mediated Isothermal Amplification Assay Combined with Enrichment Culture for Rapid Detection of Very Low Numbers of <i>Vibrio parahaemolyticus</i> in Seafood Samples

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The aim of this study was to develop and evaluate a rapid and effective method to detect Vibrio parahaemolyticus, a leading pathogen causing seafood-borne gastroenteritis. A newly designed loop-mediated isothermal amplification (LAMP) assay including a short enrichment period was optimized. This assay correctly detected all the target strains (n=61) but none of the non-target strains (n=34). Very low numbers of V. parahaemolyticus (2 colony forming unit (CFU) per gram of seafood) could be detected within 3 h and the minimum time of the whole assay was only 5 h. Comparative screening of various seafood samples (n=70) indicated that the LAMP assay is superior to polymerase chain reaction (PCR) and conventional culture methods because it is more rapid and less complex. This highly sensitive LAMP assay can be applicable as the method of choice in large-scale and rapid screening of seafood and environmental samples to detect V. parahaemolyticus strains.

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