This Article
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 Ferrándiz, M. J.
Right arrow Articles by de la Campa, A. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ferrándiz, M. J.
Right arrow Articles by de la Campa, A. G.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, November 2005, p. 4811-4813, Vol. 49, No. 11
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.11.4811-4813.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

LETTERS TO THE EDITOR

Clinical Isolates of the Spain14-5 Clone of Streptococcus pneumoniae Carry a Recombinant rpoB Gene


arrow
LETTER
 
A characterization of 61 rifampin (RIF)-resistant Streptococcus pneumoniae isolates revealed two isolates (Rif-15 and Rif-16) of the Spain14-5 clone with identical recombinant rpoB genes (5) that were collected at the Hospital Donostia (HD) from two human immunodeficiency virus-infected patients. Given that the Spain14-5 clone has been extensively studied in HD (10, 11), we have analyzed in this work 10 RIF-susceptible isolates representative of each pbp2b restriction fragment length polymorphism allelic profile among 93 isolates of the Spain14-5 clone from the HD (Table 1). The MICs, pulsed-field gel electrophoresis patterns, and rpoB sequences were determined as previously described (5). The two RIF-resistant strains carried, in addition to the H499N change responsible for RIF resistance (5), several residue changes common to five RIF-susceptible isolates (Fig. 1) and to the Streptococcus mitis NCTC 12261 type strain (5), changes that are therefore not involved in RIF resistance. Comparisons of the rpoB sequences of the 12 strains with that of S. pneumoniae R6 (Fig. 1) revealed three types of isolates: four isolates that were nonrecombinant (0.15% variation) whose sequences were identical to that of the type strain, seven recombinant isolates (R2 and R3; 4.1% variation) that had identical sequences (excluding the mutation responsible for the H499N change), and one recombinant isolate (R1; 1.2% variation) that featured two blocks of divergence, the first (L42 to T472) being identical to the sequence of S. pneumoniae R6 and the second (A473 to T700) being identical to that of strains R2 and R3.


View this table:
[in this window]
[in a new window]
  		TABLE 1. Characteristics of S. pneumoniae isolates of the Spain14-5 clone from the Hospital Donostia



View larger version (23K):
[in this window]
[in a new window]
  		FIG. 1. Genetic structure of the rpoB genes of isolates of the Spain14-5 clone. Schematic representation of RpoB showing clusters N, I, and II (A). The PCR fragments used for sequencing (gray box) and primers (black arrows) are indicated above the schematic. Nucleotide (B) and amino acid (C) sequence variations in the RpoB L42-to-T700 region of Spain14-5 isolates are also shown. The nucleotides and amino acids present at each polymorphic site are shown in full for strain R6, but for the other isolates, only sites that differ from those of R6 are shown. Codon numbers are indicated vertically above the sequences. Positions 1, 2, and 3 in the fourth row refer to the first, second, and third nucleotides in the codon, respectively. Sense mutations and amino acid changes involved in RIF resistance are shown in bold and underlined.

Recombination events between S. pneumoniae and the viri- dans streptococci of the mitis group (VSM), which have presumably occurred in the face of antibiotic selection pressure and have led to the acquisition of resistance, have been described for the genes encoding the targets of penicillin (7), fluoroquinolones (1, 4), and rifampin (5). Furthermore, recombination in genes not involved in antimicrobial resistance and therefore not subjected to selective pressure has also been detected (3, 8, 9). Given the great genetic diversity of neutral VSM genes (12), fluoroquinolone target genes (1, 6), and rpoB (M. J. Ferrándiz et al., unpublished), the most plausible explanation for the identity of the sequences of the rpoB recombinant isolates is that they derive from an ancestral isolate that underwent recombination with an RIF-susceptible VSM at, or before, 1987. Isolate Rif-16 would have acquired RIF resistance by point mutation after recombination in the presence of RIF selection (this patient had a Mycobacterium avium-disseminated infection and was treated with RIF). Transmission among patients infected with Rif-16 and Rif-15 could have occurred, since there is no evidence of RIF treatment in the patient infected with Rif-15 and both patients were admitted in the same unit of HD. However, in other cases, RIF resistance could be acquired directly by recombination (5), given that, under the ideal laboratory conditions, the frequency of transformation is several orders of magnitude greater than that of spontaneous mutation, being 5 x 10–9 (2; our unpublished results), and the frequencies of transformation are 1 x 10–2 (5) and 2 x 10–4 for chromosomal DNAs from S. pneumoniae and VSM, respectively (data not shown). The frequency of transformation in the natural environment is presumably much less than under laboratory conditions, given the low proportion of recombinants among RIF-resistant isolates (5).


arrow
REFERENCES
 
    1
  1. Balsalobre, L., M. J. Ferrándiz, J. Liñares, F. Tubau, and A. G. de la Campa. 2003. Viridans group streptococci are donors in horizontal transfer of topoisomerase IV genes to Streptococcus pneumoniae. Antimicrob. Agents Chemother. 47:2072-2081.[Abstract/Free Full Text]
    2. 2
  2. Belanger, A. E., A. Lai, M. A. Brackman, and D. J. LeBlanc. 2002. PCR-based ordered genomic libraries: a new approach to drug target identification for Streptococcus pneumoniae. Antimicrob. Agents Chemother. 46:2507-2512.[Abstract/Free Full Text]
    3. 3
  3. Coffey, T. J., M. C. Enright, M. Daniels, J. K. Morona, R. Morona, W. Hryniewicz, J. C. Paton, and B. G. Spratt. 1998. Recombinational exchanges at the capsular polysaccharide biosynthetic locus leading to frequent serotype changes among natural isolates of Streptococcus pneumoniae. Mol. Microbiol. 27:73-83.[CrossRef][Medline]
    4. 4
  4. de la Campa, A. G., L. Balsalobre, C. Ardanuy, A. Fenoll, E. Pérez-Trallero, and J. Liñares. 2004. Fluoroquinolone resistance in penicillin-resistant Streptococcus pneumoniae clones, Spain. Emerg. Infect. Dis. 10:1751-1759.[Medline]
    5. 5
  5. Ferrándiz, M., C. Ardanuy, J. Liñares, J. García-Arenzana, E. Cercenado, A. Fleites, A. G. de la Campa, and the Spanish Pneumococcal Infection Study Network. 2005. New mutations and horizontal transfer of rpoB among rifampin-resistant Streptococcus pneumoniae from four Spanish hospitals. Antimicrob. Agents Chemother. 49:2237-2245.[Abstract/Free Full Text]
    6. 6
  6. González, I., M. Georgiou, F. Alcaide, D. Balas, J. Liñares, and A. G. de la Campa. 1998. Fluoroquinolone resistance mutations in the parC, parE, and gyrA genes of clinical isolates of viridans group streptococci. Antimicrob. Agents Chemother. 42:2792-2798.[Abstract/Free Full Text]
    7. 7
  7. Hakenbeck, R., K. Kaminski, A. Köning, M. Van der Linden, J. Paik, P. Reichmann, and D. Zähner. 2000. Penicillin-binding proteins in ß-lactam-resistant Streptococcus pneumoniae. In A. Tomasz (ed.), Streptococcus pneumoniae: molecular biology and mechanism of disease. Mary Ann Liebert, Inc., Larchmont, N.Y.
    8. 8
  8. Havarstein, L. S., R. Hakenbeck, and P. Gaustad. 1997. Natural competence in the genus Streptococcus: evidence that streptococci can change pherotype by interspecies recombinational exchanges. J. Bacteriol. 179:6589-6594.[Abstract/Free Full Text]
    9. 9
  9. Martín-Galiano, A. J., L. Balsalobre, A. Fenoll, and A. G. de la Campa. 2003. Genetic characterization of optochin-susceptible viridans group streptococci. Antimicrob. Agents Chemother. 47:3187-3194.[Abstract/Free Full Text]
    10. 10
  10. Pérez-Trallero, E., J. M. Marimón, A. González, and L. Iglesias. 2003. Spain14-5 international multiresistant Streptococcus pneumoniae clone resistant to fluoroquinolones and other families of antibiotics. J. Antimicrob. Chemother. 51:715-719.[Abstract/Free Full Text]
    11. 11
  11. Pérez-Trallero, E., J. M. Marimón, A. González, D. Vicente, and J. M. García-Arenzana. 2004. Spectrum of antibiotic resistance of the Spain14-5 Streptococcus pneumoniae clone over a 22 year period. J. Antimicrob. Chemother. 53:620-625.[Abstract/Free Full Text]
    12. 12
  12. Whatmore, A. M., A. Efstratiou, A. P. Pickerill, K. Broughton, G. Woodard, D. Sturgeon, R. George, and C. G. Dowson. 2000. Genetic relationships between clinical isolates of Streptococcus pneumoniae, Streptococcus oralis, and Streptococcus mitis: characterization of "atypical" pneumococci and organisms allied to S. mitis harboring S. pneumoniae virulence factor-encoding genes. Infect. Immun. 68:1374-1382.[Abstract/Free Full Text]
M. J. Ferrándiz
Centro Nacional de Microbiología
Instituto de Salud Carlos III
Majadahonda, Madrid, Spain

E. Pérez-Trallero
Hospital Donostia
Gipuzkoa, Spain

J. M. Marimón
Hospital Donostia
Gipuzkoa, Spain

A. G. de la Campa*
Centro Nacional de Microbiología
Instituto de Salud Carlos III
Majadahonda, Madrid, Spain

* Phone: (34) 91-5097057, Fax: (34) 91-5097919, E-mail: agcampa{at}isciii.es

Antimicrobial Agents and Chemotherapy, November 2005, p. 4811-4813, Vol. 49, No. 11
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.11.4811-4813.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Marimon, J. M., Perez-Trallero, E., Ercibengoa, M., Gonzalez, A., Fenoll, A., on behalf of the Spanish Pneumococcal Infection St, (2006). Molecular epidemiology and variants of the multidrug-resistant Streptococcus pneumoniae Spain14-5 international clone among Spanish clinical isolates. J Antimicrob Chemother 57: 654-660 [Abstract] [Full Text]  

This Article
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 Ferrándiz, M. J.
Right arrow Articles by de la Campa, A. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ferrándiz, M. J.
Right arrow Articles by de la Campa, A. G.

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