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 Drancourt, M.
Right arrow Articles by Raoult, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Drancourt, M.
Right arrow Articles by Raoult, D.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, July 2001, p. 2425-2430, Vol. 39, No. 7
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.7.2425-2430.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

rpoB Sequence Analysis of Cultured Tropheryma whippelii

Michel Drancourt, Antoine Carlioz, and Didier Raoult*

Unité des Rickettsies, CNRS UPRES-A 6020, Faculté de Médecine, Université de la Méditerranée, Marseille, France

Received 8 January 2001/Returned for modification 11 March 2001/Accepted 8 April 2001

Until recently no isolate of Tropheryma whippelii was available, and therefore genetic studies were limited to those based on PCR amplification of conserved genes. In this study we determined the nucleotide sequence of rpoB (encoding the beta -subunit of RNA polymerase) from a cultured strain of T. whippelii using degenerate consensus PCR and genome walking. The T. whippelii rpoB consists of 3,657 bp with a 50.4% GC content and encodes 1,218 amino acids with a calculated molecular mass of 138 kDa. Comparison of T. whippelii RpoB with other eubacterial RpoB proteins indicated sequence similarity ranging from 57.19 (Mycoplasma pneumoniae) to 74.63% (Mycobacterium tuberculosis). Phylogenetic analysis of T. whippelii based on comparison of its RpoB sequence with sequences available for other bacteria was consistent with that previously derived from the 16S ribosomal DNA (rDNA) sequence, indicating that it belongs to the actinomyces clade. The sequence comparison allowed the design of a primer pair, TwrpoB.F and TwrpoB.R, specific for T. whippelii rpoB. When incorporated into a PCR, this primer pair allowed the detection of T. whippelii rpoB in three of three 16S rDNA PCR-positive biopsy specimens and zero of seven negative controls. rpoB could therefore be targeted in PCR-mediated detection and identification of this emerging bacterial species. This approach has previously been shown useful for the identification of related mycobacteria. This study underscores that a method involving isolation and then propagation of emerging bacteria is a useful way to quickly achieve extensive molecular knowledge of these pathogens.

* Corresponding author. Mailing address: Unité des Rickettsies, Faculté de Médecine, 27 Boulevard Jean Moulin, 13385 Marseille cedex 5, France. Phone: 33 (0)4 91 32 43 75. Fax: 33 (0)4 91 38 77 72. E-mail: Didier.Raoult{at}medecine.univ-mrs.fr.

Journal of Clinical Microbiology, July 2001, p. 2425-2430, Vol. 39, No. 7
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.7.2425-2430.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Li, W., Fenollar, F., Rolain, J.-M., Fournier, P.-E., Feurle, G. E., Muller, C., Moos, V., Marth, T., Altwegg, M., Calligaris-Maibach, R. C., Schneider, T., Biagi, F., La Scola, B., Raoult, D. (2008). Genotyping reveals a wide heterogeneity of Tropheryma whipplei. Microbiology 154: 521-527 [Abstract] [Full Text]  
  • Crapoulet, N., Robineau, S., Raoult, D., Renesto, P. (2005). Intervening Sequence Acquired by Lateral Gene Transfer in Tropheryma whipplei Results in 23S rRNA Fragmentation. Appl. Environ. Microbiol. 71: 6698-6701 [Abstract] [Full Text]  
  • Sloan, L. M., Rosenblatt, J. E., Cockerill, F. R. III (2005). Detection of Tropheryma whipplei DNA in Clinical Specimens by LightCycler Real-Time PCR. J. Clin. Microbiol. 43: 3516-3518 [Abstract] [Full Text]  
  • Hurtle, W., Bode, E., Kulesh, D. A., Kaplan, R. S., Garrison, J., Bridge, D., House, M., Frye, M. S., Loveless, B., Norwood, D. (2004). Detection of the Bacillus anthracis gyrA Gene by Using a Minor Groove Binder Probe. J. Clin. Microbiol. 42: 179-185 [Abstract] [Full Text]  
  • Fenollar, F., Fournier, P.-E., Robert, C., Raoult, D. (2004). Use of Genome Selected Repeated Sequences Increases the Sensitivity of PCR Detection of Tropheryma whipplei. J. Clin. Microbiol. 42: 401-403 [Abstract] [Full Text]  
  • Raoult, D., Ogata, H., Audic, S., Robert, C., Suhre, K., Drancourt, M., Claverie, J.-M. (2003). Tropheryma whipplei Twist: A Human Pathogenic Actinobacteria With a Reduced Genome. Genome Res 13: 1800-1809 [Abstract] [Full Text]  
  • Ko, K. S., Kim, J.-M., Kim, J.-W., Jung, B. Y., Kim, W., Kim, I. J., Kook, Y.-H. (2003). Identification of Bacillus anthracis by rpoB Sequence Analysis and Multiplex PCR. J. Clin. Microbiol. 41: 2908-2914 [Abstract] [Full Text]  
  • Masselot, F., Boulos, A., Maurin, M., Rolain, J. M., Raoult, D. (2003). Molecular Evaluation of Antibiotic Susceptibility: Tropheryma whipplei Paradigm. Antimicrob. Agents Chemother. 47: 1658-1664 [Abstract] [Full Text]  
  • Fenollar, F., Fournier, P.-E., Raoult, D., Gerolami, R., Lepidi, H., Poyart, C. (2002). Quantitative Detection of Tropheryma whipplei DNA by Real-Time PCR. J. Clin. Microbiol. 40: 1119-1120 [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»