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Applied and Environmental Microbiology, October 2006, p. 6593-6599, Vol. 72, No. 10
0099-2240/06/$08.00+0     doi:10.1128/AEM.03068-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Immunoquantitative Real-Time PCR for Detection and Quantification of Staphylococcus aureus Enterotoxin B in Foods

Andreja Rajkovic,1 Benaissa El Moualij,2 Mieke Uyttendaele,1* Philippe Brolet,2 Willy Zorzi,2 Ernst Heinen,2 Ellen Foubert,1 and Johan Debevere1

Laboratory of Food Microbiology and Food Preservation, Department of Food Safety and Food Quality, Faculty of Bioscience Engineering, Ghent University, Ghent,1 Laboratory of Human Histology, University of Liege, Liege, Belgium2

Received 29 December 2005/ Accepted 31 July 2006

A real-time immunoquantitative PCR (iqPCR) method for detection of Staphylococcus aureus enterotoxin B (SEB) was developed and evaluated using both pure cultures and foods. The assay consisted of immunocapture of SEB and real-time PCR amplification of the DNA probe linked to the detection antibody. iqPCR was compared to an in-house enzyme-linked immunosorbent assay (ELISA) using the same couple of capture-detection antibodies and to commercial kits for detection of S. aureus enterotoxins (SE). The iqPCR was approximately 1,000 times more sensitive (<10 pg ml–1) than the in-house ELISA and had a dynamic range of approximately 10 pg ml–1 to approximately 30,000 pg ml–1. iqPCR was not inhibited by any of the foods tested and was able to detect SEB present in these foods. No cross-reactivity with SE other than SEB was observed. Application of iqPCR for detection of SEB in cultures of S. aureus revealed the onset of SEB production after 4 h of incubation at 22, 37, and 42°C, which was in the first half of the exponential growth phase. The total amounts of SEB produced by the two strains tested were larger at 42°C than at 37°C and were strain dependent.

* Corresponding author. Mailing address: Coupure links 653, B-9000 Ghent, Belgium. Phone: 32 9 264 61 78. Fax: 32 9 225 55 10. E-mail: mieke.uyttendaele{at}UGent.be.

Applied and Environmental Microbiology, October 2006, p. 6593-6599, Vol. 72, No. 10
0099-2240/06/$08.00+0     doi:10.1128/AEM.03068-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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