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

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.