This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by McKillip, J. L.
Right arrow Articles by Drake, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by McKillip, J. L.
Right arrow Articles by Drake, M.
Agricola
Right arrow Articles by McKillip, J. L.
Right arrow Articles by Drake, M.

 Previous Article  |  Next Article 

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:H7dagger

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.

dagger 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.


This article has been cited by other articles:

  • Matsuda, K., Tsuji, H., Asahara, T., Kado, Y., Nomoto, K. (2007). Sensitive Quantitative Detection of Commensal Bacteria by rRNA-Targeted Reverse Transcription-PCR. Appl. Environ. Microbiol. 73: 32-39 [Abstract] [Full Text]  
  • Pecson, B. M., Barrios, J. A., Johnson, D. R., Nelson, K. L. (2006). A Real-Time PCR Method for Quantifying Viable Ascaris Eggs Using the First Internally Transcribed Spacer Region of Ribosomal DNA. Appl. Environ. Microbiol. 72: 7864-7872 [Abstract] [Full Text]  
  • Aellen, S., Que, Y.-A., Guignard, B., Haenni, M., Moreillon, P. (2006). Detection of Live and Antibiotic-Killed Bacteria by Quantitative Real-Time PCR of Specific Fragments of rRNA.. Antimicrob. Agents Chemother. 50: 1913-1920 [Abstract] [Full Text]  
  • Nocker, A., Camper, A. K. (2006). Selective removal of DNA from dead cells of mixed bacterial communities by use of ethidium monoazide.. Appl. Environ. Microbiol. 72: 1997-2004 [Abstract] [Full Text]  
  • Suenaga, H., Liu, R., Shiramasa, Y., Kanagawa, T. (2005). Novel Approach to Quantitative Detection of Specific rRNA in a Microbial Community, Using Catalytic DNA. Appl. Environ. Microbiol. 71: 4879-4884 [Abstract] [Full Text]  
  • Rudi, K., Moen, B., Dromtorp, S. M., Holck, A. L. (2005). Use of Ethidium Monoazide and PCR in Combination for Quantification of Viable and Dead Cells in Complex Samples. Appl. Environ. Microbiol. 71: 1018-1024 [Abstract] [Full Text]  
  • Dreier, J., Stormer, M., Kleesiek, K. (2004). Two Novel Real-Time Reverse Transcriptase PCR Assays for Rapid Detection of Bacterial Contamination in Platelet Concentrates. J. Clin. Microbiol. 42: 4759-4764 [Abstract] [Full Text]  
  • McIngvale, S. C., Elhanafi, D., Drake, M. A. (2002). Optimization of Reverse Transcriptase PCR To Detect Viable Shiga-Toxin-Producing Escherichia coli. Appl. Environ. Microbiol. 68: 799-806 [Abstract] [Full Text]  
  • Li, W., Drake, M. A. (2001). Development of a Quantitative Competitive PCR Assay for Detection and Quantification of Escherichia coli O157:H7 Cells. Appl. Environ. Microbiol. 67: 3291-3294 [Abstract] [Full Text]  
  • Hu, Y., Mangan, J. A., Dhillon, J., Sole, K. M., Mitchison, D. A., Butcher, P. D., Coates, A. R. M. (2000). Detection of mRNA Transcripts and Active Transcription in Persistent Mycobacterium tuberculosis Induced by Exposure to Rifampin or Pyrazinamide. J. Bacteriol. 182: 6358-6365 [Abstract] [Full Text]  
  • Nogva, H. K., Bergh, A., Holck, A., Rudi, K. (2000). Application of the 5'-Nuclease PCR Assay in Evaluation and Development of Methods for Quantitative Detection of Campylobacter jejuni. Appl. Environ. Microbiol. 66: 4029-4036 [Abstract] [Full Text]  
  • Lucore, L. A., Cullison, M. A., Jaykus, L.-A. (2000). Immobilization with Metal Hydroxides as a Means To Concentrate Food-Borne Bacteria for Detection by Cultural and Molecular Methods. Appl. Environ. Microbiol. 66: 1769-1776 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»