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Applied and Environmental Microbiology, March 2007, p. 1892-1898, Vol. 73, No. 6
0099-2240/07/$08.00+0     doi:10.1128/AEM.02219-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Diagnostic Real-Time PCR Assays for the Detection of Emetic Bacillus cereus Strains in Foods and Recent Food-Borne Outbreaks{triangledown} ,{dagger}

Martina Fricker,1 Ute Messelhäußer,2 Ulrich Busch,2 Siegfried Scherer,1 and Monika Ehling-Schulz1*

Abteilung Mikrobiologie, Zentralinstitut für Ernährungs- und Lebensmittelforschung (ZIEL), Technische Universität München, D-85354 Freising, Germany,1 Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit (LGL), D-85764 Oberschleißheim, Germany2

Received 21 September 2006/ Accepted 18 January 2007

Cereulide-producing Bacillus cereus can cause an emetic type of food-borne disease that mimics the symptoms provoked by Staphylococcus aureus. Based on the recently discovered genetic background for cereulide formation, a novel 5' nuclease (TaqMan) real-time PCR assay was developed to provide a rapid and sensitive method for the specific detection of emetic B. cereus in food. The TaqMan assay includes an internal amplification control and primers and a probe designed to target a highly specific part of the cereulide synthetase genes. Additionally, a specific SYBR green I assay was developed and extended to create a duplex SYBR green I assay for the one-step identification and discrimination of the two emesis-causing food pathogens B. cereus and S. aureus. The inclusivity and exclusivity of the assay were assessed using a panel of 100 strains, including 23 emetic B. cereus and 14 S. aureus strains. Different methods for DNA isolation from artificially contaminated foods were evaluated, and established real-time assays were used to analyze two recent emetic food poisonings in southern Germany. One of the food-borne outbreaks included 17 children visiting a day care center who vomited after consuming a reheated rice dish, collapsed, and were hospitalized; the other case concerned a single food-poisoning incident occurring after consumption of cauliflower. Within 2 h, the etiological agent of these food poisonings was identified as emetic B. cereus by using the real-time PCR assay.

* Corresponding author. Mailing address: Abt. Mikrobiologie/ZIEL, TUM, Weihenstephaner Berg 3, 85354 Freising, Germany. Phone: 49-8161-713851. Fax: 49-8161-714492. E-mail: monika.ehling-schulz{at}wzw.tum.de.

{triangledown} Published ahead of print on 26 January 2007.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, March 2007, p. 1892-1898, Vol. 73, No. 6
0099-2240/07/$08.00+0     doi:10.1128/AEM.02219-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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