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Lett Appl Microbiol. 2008 Jun;46(6):649-54. doi: 10.1111/j.1472-765X.2008.02368.x. Epub 2008 Apr 17.

The development of rapid real-time PCR detection system for Vibrio parahaemolyticus in raw oyster.

Author information

  • 1Food Research & Development Center, Samsung Everland Inc., Mabuk-dong, Giheung-gu, Yongin, South Korea. jeongsoon.kim@samsung.com

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 microl of de-ionized water. DNA was extracted by boiling for 20 min, and 0.5 microl was used as a template for PCR reaction. Real-time PCR was performed with TMC-1000 system (1 microl 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.

PMID:
18422939
[PubMed - indexed for MEDLINE]
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