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| Rapid Detection of Vibrio parahaemolyticus in Seafood Using Loop-Mediated Isothermal Amplification (LAMP) Method |
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Abstract Vibrio parahaemolyticus has been considered as one of the most important foodborne bacterial pathogens. In this paper, the loop-mediated isothermal amplification (LAMP) that amplifies DNA with high specificity and rapidity under an isothermal condition was applied for rapid detection of this pathogen for the first time. A set of four primers, two outer and two inner primers, was designed specifically to recognize the thermolabile hemolysin gene (tlh) of V. parahaemolyticus in this study. Genomic DNAs from 28 bacterial strains including 14 V. parahaemolyticus strains were amplified using LAMP, and no amplicon was observed in other bacterial strains. The detection limit of this LAMP assay was around 90 fg/LAMP Mixture of V.parahaemolyticus genomic DNA, 24 colony forming units for pure cultures, 89 cfu/g for non-cultured artificially contaminated food samples. In addition, 40 seafood samples were tested, and 8 samples were found V. parahaemolyticus positive by LAMP. Among the tested positive samples, 6 samples were detected positive by traditional culture methods. These results suggested that detection of V. parahaemolyticus by LAMP is an effective and low-cost procedure with high specificity and sensitivity that requires no specialized equipment. This assay is expected to become a valuable tool for rapid detection and identification of V. parahaemolyticus.
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Received: 25 December 2007
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