Food Science and Biotechnology

Korean Society of Food Science and Technology (한국식품과학회)
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A Multiplex PCR Assay for the Detection and Differentiation of Enterotoxin-producing and Emetic Toxin-producing Bacillus cereus Strains

  • Lee, Dae-Sung (Department of Food Science and Technology, Dongguk University) ;
  • Kim, Keun-Sung (Department of Food Science and Technology, Chung-Ang University) ;
  • Kwon, Ki-Sung (Department of Food Microbiology, Korea Food & Drug Administration) ;
  • Hong, Kwang-Won (Department of Food Science and Technology, Dongguk University)
  • Published : 2008.08.31
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Abstract

Bacillus cereus causes two different types of food poisoning syndromes: diarrhea and emesis. The diarrheal syndrome is attributed to various enterotoxins, including nonhemolytic enterotoxin, hemolytic enterotoxin, and enterotoxin-T, whereas the emetic syndrome is caused by the dodecadepsipeptide toxin cereulide. A multiplex polymerase chain reaction (PCR) assay was developed to rapidly detect and identify B. cereus strains. Three primer pairs specific to regions within genes encoding nonhemolytic enterotoxin (nheA), molecular chaperonin (groEL), and cereulide synthetase (ces) were used to identify and differentiate between the enterotoxin-producing and emetic toxin-producing B. cereus strains. The cereulide-producing emetic B. cereus showed 3 PCR products of 325, 405, and 685 bp for the groEL, ces, and nheA genes, respectively, whereas the enterotoxin-producing B. cereus showed 2 PCR products without a ces gene specific DNA fragment. Specific amplifications and differentiations by multiplex PCR assay were obtained using 62 B. cereus strains and 13 strains' of other bacterial species. The detection limit of this assay for enterotoxin-producing strain and emetic toxin-producing strain from pure cultures were $2.4{\times}10^1$ and $6.0{\times}10^2\;CFU/tube$, respectively. These results suggest that our multiplex PCR method may be useful for the rapid detection and differentiation of B. cereus strains in foods.

Keywords

References

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