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Appl Environ Microbiol, May 1998, p. 1640-1643, Vol. 64, No. 5
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
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
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.
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