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Applied and Environmental Microbiology, November 1998, p. 4264-4268, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
rRNA Stability in Heat-Killed and UV-Irradiated Enterotoxigenic
Staphylococcus aureus and Escherichia coli
O157:H7
John L.
McKillip,1
Lee-Ann
Jaykus,2 and
Maryanne
Drake1,*
Department of Food Science and Technology,
Southeast Dairy Foods Research Center, Mississippi State University,
Mississippi State, Mississippi 39762-9805,1 and
Department of Food Science, Southeast Dairy Foods Research
Center, North Carolina State University, Raleigh, North Carolina
27695-76242
Received 11 June 1998/Accepted 7 August 1998
Differentiation of viable cells from nonviable cells is of
considerable importance in the development of methods to detect foodborne pathogens. To study the suitability of 16S rRNA as an indicator of cell viability in nucleic acid-based detection assays, we
examined rRNA stability in two representative foodborne pathogens, Escherichia coli O157:H7 and enterotoxigenic
Staphylococcus aureus, which were inactivated by extreme
heat, moderate heat, and UV irradiation. Cell death under all
conditions was confirmed by a failure to grow in brain heart infusion
broth after incubation for 48 h at 37°C. rRNA stability was
monitored by a Northern blot analysis, and detection was evaluated by
using reverse transcription (RT)-PCR performed with two primer sets
(which produced 325- and 1,400-bp amplicons). rRNA of neither pathogen
was detected by Northern blot analysis and RT-PCR after cells were
killed by autoclaving at 121°C for 15 min. In contrast, intact rRNA
of both pathogens were detected by Northern blotting and could be
amplified by RT-PCR up to 48 h after cells were killed by heat
treatment at 80°C and UV irradiation at 254 nm. rRNA was a suitable
target molecule for monitoring bacterial viability under extreme heat
conditions, but the presence of rRNA was not correlated with viability
following moderate heat inactivation or UV irradiation of cells.
*
Corresponding author. Mailing address: Department of
Food Science and Technology, Southeast Dairy Foods Research Center, Box 9805, Mississippi State University, Mississippi State, MS 39762-9805. Phone: (601) 325-3200. Fax: (601) 325-8728. E-mail:
maryanne{at}ra.msstate.edu.

This is Mississippi Agricultural and Forestry Experiment Station
project number
J9387.
Applied and Environmental Microbiology, November 1998, p. 4264-4268, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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