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JAC Advance Access published online on November 24, 2004
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkh481
© 2004 by The British Society for Antimicrobial Chemotherapy
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Received August 5, 2004
Revised September 24, 2004
Accepted September 28, 2004

Original article

Distribution and molecular analysis of mef(A)-containing elements in tetracycline-susceptible and -resistant Streptococcus pyogenes clinical isolates with efflux-mediated erythromycin resistance

Andrea Brenciani 1, Kayode K. Ojo 2, Alessia Monachetti 1, Stefano Menzo 1, Marilyn C. Roberts 2, Pietro E. Varaldo 1*, and Eleonora Giovanetti 1

1 Institute of Microbiology and Biomedical Sciences, Marche Polytechnic University, 60131 Ancona, Italy
2 Department of Pathobiology, University of Washington, Seattle, WA 98195-7238, USA

* To whom correspondence should be addressed.
Pietro E. Varaldo, E-mail: pe.varaldo{at}univpm.it


  Abstract

Objectives: To analyse the distribution and molecular features of mef(A)-containing elements in a large collection of different Streptococcus pyogenes clinical isolates with efflux-mediated erythromycin resistance. To further characterize a tet(O)-mef(A) element.

Methods: Gene detection was carried out by PCR using primers designed from established sequences or from sequences in this study. From a tet(O)-mef(A) element (~60 kb), an 11 972 bp region including the tet(O) and mef(A) genes was sequenced.

Results: In the tetracycline-susceptible isolates (n =28), the mef(A) gene was contained in a regular Tn1207.1 transposon (7.2 kb), which was inserted into one of two previously described elements, Tn1207.3 (~52 kb) or a 58.8 kb chimeric element, both flanked by the comEC gene. In the tetracycline-resistant isolates (n =61), all of which carried the tet(O) gene, the mef(A) gene was part of a variable Tn1207.1-related transposon inserted into unique elements which contained the tet(O) gene ~2.3 to 5.5 kb upstream of the mef(A) gene and were not flanked by the comEC gene. In the Tn1207.1-like transposon of these tet(O)-mef(A) elements, only msr(D) (orf5) and a modified orf6, in addition to mef(A), were detected by PCR in all isolates tested; while orf1 and orf2 were always undetectable, orf3, orf7 and orf8 were found in variable percentages. In an orf3-positive element, sequencing identified four new open reading frames downstream of the tet(O) gene, followed by three short sequences with homology to sequences of the pneumococcal mega element.

Conclusions: The mef(A) gene is carried on different chromosomal genetic elements depending on whether the isolates are susceptible or resistant to tetracycline.

Keywords: tet(O) gene; Tn1207.1 transposon; macrolides; M phenotype; chimeric elements.
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