The development of rapid real-time PCR detection system for Vibrio parahaemolyticus in raw oyster |
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Authors: | Kim J S Lee G G Kim J Kwon J Y Kwon S-T |
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Affiliation: | Food Research &Development Center, Samsung Everland Inc., Mabuk-dong, Giheung-gu, Yongin, South Korea; Department of Genetic Engineering, Sungkyunkwan University, Chunchon-dong, Jangan-gu, Suwon, South Korea |
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Abstract: | 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 μ l of de-ionized water. DNA was extracted by boiling for 20 min, and 0·5 μ l was used as a template for PCR reaction. Real-time PCR was performed with TMC-1000 system (1 μ l 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. |
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Keywords: | Vibrio parahaemolyticus micro PCR TMC-1000 PCR system |
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