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 Rousseaux, S.
Right arrow Articles by Guzzo, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Rousseaux, S.
Right arrow Articles by Guzzo, J.
Agricola
Right arrow Articles by Rousseaux, S.
Right arrow Articles by Guzzo, J.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, April 2004, p. 2180-2185, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.2180-2185.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Use of PCR-Restriction Fragment Length Polymorphism of inlA for Rapid Screening of Listeria monocytogenes Strains Deficient in the Ability To Invade Caco-2 Cells

S. Rousseaux, M. Olier, J. P. Lemaître, P. Piveteau, and J. Guzzo*

Laboratoire de Microbiologie, UMR INRA 1232, ENSBANA, 21000 Dijon, France

Received 9 June 2003/ Accepted 5 January 2004

A PCR-restriction fragment length polymorphism (RFLP) method was developed in order to screen a large number of strains for impaired adhesion to epithelial cells due to expression of truncated InlA. inlA polymorphism was analyzed by PCR-RFLP in order to correlate inlA PCR-RFLP profiles and production of truncated InlA. Thirty-seven Listeria monocytogenes strains isolated from various sources, including five noninvasive and two invasive reference strains, were screened. Two endonucleases (AluI and Tsp509I) were used, and they generated five composite profiles. Thirteen L. monocytogenes isolates were characterized by two specific PCR-RFLP profiles similar to PCR-RFLP profiles of noninvasive reference strains previously described as strains that produce truncated InlA. Ten of the 13 isolates showed low abilities to invade human epithelial Caco-2 cells. However, 4 of the 13 isolates were able to invade Caco-2 cells like reference strains containing complete InlA. Sequencing of inlA and Western blot analysis confirmed that truncated InlA was expressed in the 10 L. monocytogenes strains which were isolated from food. This PCR-RFLP method allowed us to identify 10 new strains expressing a truncated internalin. Based on the results obtained in this study, the PCR-RFLP method seems to be an interesting method for rapidly screening L. monocytogenes strains deficient in the ability to invade Caco-2 cells when a sizeable number of strains are studied.

* Corresponding author. Mailing address: Laboratoire de Microbiologie, UMR INRA 1232, ENSBANA, 1 Esplanade Erasme, 21000 Dijon, France. Phone: 33 3 80 39 66 75. Fax: 3 3 80 39 39 55. E-mail: jguzzo{at}u-bourgogne.fr.

Applied and Environmental Microbiology, April 2004, p. 2180-2185, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.2180-2185.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Ward, T. J., Ducey, T. F., Usgaard, T., Dunn, K. A., Bielawski, J. P. (2008). Multilocus Genotyping Assays for Single Nucleotide Polymorphism-Based Subtyping of Listeria monocytogenes Isolates. Appl. Environ. Microbiol. 74: 7629-7642 [Abstract] [Full Text]  
  • Van Stelten, A., Nightingale, K. K. (2008). Development and Implementation of a Multiplex Single-Nucleotide Polymorphism Genotyping Assay for Detection of Virulence-Attenuating Mutations in the Listeria monocytogenes Virulence-Associated Gene inlA. Appl. Environ. Microbiol. 74: 7365-7375 [Abstract] [Full Text]  
  • Nightingale, K. K., Ivy, R. A., Ho, A. J., Fortes, E. D., Njaa, B. L., Peters, R. M., Wiedmann, M. (2008). inlA Premature Stop Codons Are Common among Listeria monocytogenes Isolates from Foods and Yield Virulence-Attenuated Strains That Confer Protection against Fully Virulent Strains. Appl. Environ. Microbiol. 74: 6570-6583 [Abstract] [Full Text]  
  • Lyautey, E., Lapen, D. R., Wilkes, G., McCleary, K., Pagotto, F., Tyler, K., Hartmann, A., Piveteau, P., Rieu, A., Robertson, W. J., Medeiros, D. T., Edge, T. A., Gannon, V., Topp, E. (2007). Distribution and Characteristics of Listeria monocytogenes Isolates from Surface Waters of the South Nation River Watershed, Ontario, Canada. Appl. Environ. Microbiol. 73: 5401-5410 [Abstract] [Full Text]  
  • Orsi, R. H., Ripoll, D. R., Yeung, M., Nightingale, K. K., Wiedmann, M. (2007). Recombination and positive selection contribute to evolution of Listeria monocytogenes inlA. Microbiology 153: 2666-2678 [Abstract] [Full Text]  
  • Felicio, M. T. S., Hogg, T., Gibbs, P., Teixeira, P., Wiedmann, M. (2007). Recurrent and Sporadic Listeria monocytogenes Contamination in Alheiras Represents Considerable Diversity, Including Virulence-Attenuated Isolates. Appl. Environ. Microbiol. 73: 3887-3895 [Abstract] [Full Text]  
  • Nightingale, K. K., Lyles, K., Ayodele, M., Jalan, P., Nielsen, R., Wiedmann, M. (2006). Novel Method To Identify Source-Associated Phylogenetic Clustering Shows that Listeria monocytogenes Includes Niche-Adapted Clonal Groups with Distinct Ecological Preferences.. J. Clin. Microbiol. 44: 3742-3751 [Abstract] [Full Text]  
  • Werbrouck, H., Grijspeerdt, K., Botteldoorn, N., Van Pamel, E., Rijpens, N., Van Damme, J., Uyttendaele, M., Herman, L., Van Coillie, E. (2006). Differential inlA and inlB Expression and Interaction with Human Intestinal and Liver Cells by Listeria monocytogenes Strains of Different Origins.. Appl. Environ. Microbiol. 72: 3862-3871 [Abstract] [Full Text]  
  • Nightingale, K. K., Windham, K., Martin, K. E., Yeung, M., Wiedmann, M. (2005). Select Listeria monocytogenes Subtypes Commonly Found in Foods Carry Distinct Nonsense Mutations in inlA, Leading to Expression of Truncated and Secreted Internalin A, and Are Associated with a Reduced Invasion Phenotype for Human Intestinal Epithelial Cells. Appl. Environ. Microbiol. 71: 8764-8772 [Abstract] [Full Text]  
  • Olier, M., Garmyn, D., Rousseaux, S., Lemaitre, J.-P., Piveteau, P., Guzzo, J. (2005). Truncated Internalin A and Asymptomatic Listeria monocytogenes Carriage: In Vivo Investigation by Allelic Exchange. Infect. Immun. 73: 644-648 [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»