• Vibrio parahaemolyticus;
  • micro PCR;
  • TMC-1000 PCR system


Aims:  To develop a new rapid real-time polymerase chain reaction (PCR) based detection system for Vibrio parahaemolyticus (V. parahaemolyticus) applicable to raw oyster samples.

Methods and Results: V. parahaemolyticus cells were artificially inoculated to oysters. Samples were homogenized in 100 ml of sterile saline water and serially diluted to 1·5 CFU ml−1 level. One millilitre of diluents was centrifuged and the pellet was resuspended with 100 μl of de-ionized water. DNA was extracted by boiling for 20 min, and 0·5 μl was used as a template for PCR reaction. Real-time PCR was performed with TMC-1000 system (1 μl PCR system). The detection system was found to achieve detection limit of 1·5 CFU g−1 for V. parahaemolyticus. Furthermore, the specificities of these assay systems were confirmed with more than 20 bacterial strains, including various Vibrio species.

Conclusions:  Rapid and sensitive food-borne pathogen detection techniques for V. parahaemolyticus is important to the food industry and consumers. The direct detection of V. parahaemolyticus from food is possible with micro real-time PCR system.

Significance and Impact of the Study:  This study shows that oyster samples can be tested for V. parahaemolyticus with a rapid, specific and simple procedure.