Food Science and Biotechnology

Korean Society of Food Science and Technology (한국식품과학회)
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Improved Detection of ${\gamma}-Irradiated$ Vibrio vulnificus after Heat and Cold Shock Treatment by Using Ethidium Monoazide Real-time PCR

  • Lee, Jung-Lim (Department of Food Science, Massachusetts Agricultural Experiment Station, University of Massachusetts) ;
  • Levin, Robert E. (Department of Food Science, Massachusetts Agricultural Experiment Station, University of Massachusetts)
  • Published : 2009.06.30
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Abstract

Gamma $({\gamma})-irradiation$ can be used to control pathogens such as Vibrio vulnificus in seafood. The effects of irradiation on microbial cell populations (%) have been studied in order to develop detection methods for irradiated foods. The method used in this study was ethidium bromide monoazide (EMA) real-time polymerase chain reaction (PCR), using V. vulnificus specific primer, EMA, and $SYBR^{(R)}$ Green to discriminate between ${\gamma}-irradiated$ and non-irradiated cells. Confocal microscope examination showed that ${\gamma}-irradiation$ damaged portions of the cell membrane, allowing EMA to penetrate cells of irradidated V. vulnificus. ${\gamma}-Irradiation$ at 1.08 KGy resulted in log reduction ($-1.15{\pm}0.13$ log reduction) in genomic targets derived from EMA real-time PCR. The combination cold/heat shock resulted in the highest ($-1.74{\pm}0.1$ log reduction) discrimination of dead irradiated V. vulnificus by EMA real-time PCR.

Keywords

References

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