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Appl Environ Microbiol, May 1998, p. 1640-1643, Vol. 64, No. 5
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Influence of Cold Stress on the Preliminary Enrichment Time Needed for Detection of Enterohemorrhagic Escherichia coli in Ground Beef by PCR

M. Uyttendaele,1 C. Grangette,2 F. Rogerie,2 S. Pasteau,2 J. Debevere,1 and M. Lange2,*

Department of Food Technology and Nutrition, Faculty of Agricultural and Applied Biological Sciences, University of Ghent, 9000 Ghent, Belgium,1 and Service de Recherche et Développement, Institut Pasteur de Lille, 59019 Lille Cedex, France2

Received 30 September 1997/Accepted 23 February 1998

The influence of cold stress at 4 and 0°C on the detection time as assessed by impedance technology (Bactometer; Biomérieux, Marcy l'Etoile, France) of different enterohemorrhagic Escherichia coli (EHEC) strains was determined. Although there is some variation in susceptibility among EHEC strains, prolonged exposure of EHEC to cold stress, i.e., 4 and 5 days at 4 and 0°C, respectively, in general significantly increased their detection time. This reflects an increase of the lag-phase time caused by cold stress. Two EHEC strains were selected to determine the minimum preliminary enrichment time that would ensure a positive PCR detection of low numbers of verotoxin-producing E. coli (VTEC; 2 to 2 × 105 CFU/25 g) inoculated into ground beef (25 g) and stored at 4 or -20°C for 8 and 14 days, respectively. Incubation times of 6 and 9 h of 1 to 10 CFU/g and 1 to 10 CFU/25 g, respectively, were sufficient for PCR detection of VTEC in ground beef when analysis was performed immediately after inoculation (no cold stress). When cells are exposed to cold stress (4 or -20°C) a 24-h enrichment period is recommended. Restriction of enrichment time to 9 h under these circumstances decreases the sensitivity of PCR detection to 80 CFU/g. Hence, to obtain maximum sensitivity, PCR detection of VTEC in naturally contaminated ground beef should be performed after 24 h of enrichment.


* Corresponding author. Mailing address: Service de Recherche et Développement, Institut Pasteur de Lille, rue de Professeur Calmette 1---B.P.245, 59019 Lille Cedex, France. Phone: 33 (03) 20 87 72 08. Fax: 33 (03) 20 87 72 06. E-mail: Marc.Lange{at}pasteur-lille.fr.


Appl Environ Microbiol, May 1998, p. 1640-1643, Vol. 64, No. 5
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.



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