Simultaneous enrichment of Salmonella spp, Escherichia coli O157:H7, Vibrio parahaemolyticus, Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes by single broth and screening of the pathogens by multiplex real-time PCR

Hiroshi Kobayashi, Jun Kubota, Kumiko Fujihara, Ken-ichi Honjoh, Masayoshi Iio, Naomi Fujiki, Mika Nakabe, Shun Ichi Oda, Toshiya Satoyama, Kazushige Takasu, Hisao Nakanishi, Takahisa Miyamoto

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Simultaneous enrichment broth (SEB) was developed for the single-enrichment simultaneous screening of six major food poisoning bacteria. After enrichment in SEB for 18 h at 37°C, viable counts of six major food poisoning bacteria (Staphylococcus aureus, Salmonella spp., Escherichia coli O157:H7, Vibrio parahaemolyticus, Bacillus cereus, and Listeria monocytogenes) were sufficient for 5' nuclease multiplex real-time PCR assay using existing primers and probes. By labeling the probes with three different fluorescent dyes, the assay could be carried out in 2 tubes. The whole process, including enrichment and PCR, was completed within 24 h and the detection limits for the target bacteria from the food sample (boiled chicken) were 36 cfu/25 g for S. aureus, 5.3 cfu/25 g for Salmonella spp, 2.9 cfu/25 g for E. coli O157:H7, 2.0 cfu/25 g for V. parahaemolyticus, 5.5 cfu/25 g for B. cereus, and 6.2 cfu/25 g for L. monocytogenes. The results were comparable to conventional methods that require 4-6 days.

Original languageEnglish
Pages (from-to)427-438
Number of pages12
JournalFood Science and Technology Research
Volume15
Issue number4
DOIs
Publication statusPublished - Jul 1 2009

Fingerprint

Bacillus cereus
Listeria
Vibrio parahaemolyticus
Salmonella
Escherichia coli O157
Multiplex Polymerase Chain Reaction
Listeria monocytogenes
Pathogens
Escherichia coli
Staphylococcus aureus
Real-Time Polymerase Chain Reaction
Bacteria
Screening
quantitative polymerase chain reaction
Foodborne Diseases
foodborne illness
screening
pathogens
Assays
bacteria

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science
  • Chemical Engineering(all)
  • Industrial and Manufacturing Engineering
  • Marketing

Cite this

Simultaneous enrichment of Salmonella spp, Escherichia coli O157:H7, Vibrio parahaemolyticus, Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes by single broth and screening of the pathogens by multiplex real-time PCR. / Kobayashi, Hiroshi; Kubota, Jun; Fujihara, Kumiko; Honjoh, Ken-ichi; Iio, Masayoshi; Fujiki, Naomi; Nakabe, Mika; Oda, Shun Ichi; Satoyama, Toshiya; Takasu, Kazushige; Nakanishi, Hisao; Miyamoto, Takahisa.

In: Food Science and Technology Research, Vol. 15, No. 4, 01.07.2009, p. 427-438.

Research output: Contribution to journalArticle

Kobayashi, Hiroshi ; Kubota, Jun ; Fujihara, Kumiko ; Honjoh, Ken-ichi ; Iio, Masayoshi ; Fujiki, Naomi ; Nakabe, Mika ; Oda, Shun Ichi ; Satoyama, Toshiya ; Takasu, Kazushige ; Nakanishi, Hisao ; Miyamoto, Takahisa. / Simultaneous enrichment of Salmonella spp, Escherichia coli O157:H7, Vibrio parahaemolyticus, Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes by single broth and screening of the pathogens by multiplex real-time PCR. In: Food Science and Technology Research. 2009 ; Vol. 15, No. 4. pp. 427-438.
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