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JAC Advance Access originally published online on February 16, 2006
Journal of Antimicrobial Chemotherapy 2006 57(4):654-660; doi:10.1093/jac/dkl028
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© The Author 2006. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Molecular epidemiology and variants of the multidrug-resistant Streptococcus pneumoniae Spain14-5 international clone among Spanish clinical isolates

José María Marimón1, Emilio Pérez-Trallero1,2,*, María Ercibengoa1, Alberto Gonzalez1, Asunción Fenoll3 on behalf of the Spanish Pneumococcal Infection Study Network (G03/103)

1 Servicio de Microbiología, Hospital Donostia Paseo Dr Beguiristain s/n 20014 San Sebastián, Spain; 2 Departamento de Medicina Preventiva y Salud Pública, Universidad del País Vasco, Paseo Dr Beguiristain s/n 20014 San Sebastián, Spain; 3 Centro Nacional de Microbiología, Majadahonda, Madrid, Spain

Received 23 November 2005; returned 5 January 2006; revised 17 January 2006; accepted 19 January 2006

* Corresponding author. Tel: +34-94-300-7046; Fax: +34-94-300-7063; E-mail: mikrobiol{at}terra.es

Objective: To analyse the molecular structure of several antimicrobial resistance determinants in isolates of the Spain14-5 clone to better understand its emergence and spread.

Methods: The distinct genes and mechanism of resistance to penicillin, erythromycin, clindamycin, tetracycline, chloramphenicol and trimethoprim were studied in an apparently homogeneous group of 117 isolates of the multidrug-resistant Spain14-5 major clone isolated in Spain between 1981 and 2004.

Results: Several genotyping techniques such as PFGE, BOX-PCR and multilocus sequence typing (MLST) revealed a high degree of homogeneity among these isolates over time. Nevertheless, distinct variants of the clone could be established according to the restriction fragment length polymorphism (RFLP) patterns of the penicillin-binding protein (pbp) genes and the sequences of the dihydrofolate reductase (dhfr) gene. In addition, an association between the pbp2b RFLP patterns, the ddl alleles identified by MLST and the dhfr alleles was found. The emergence of variants of the Spain14-5 clone, which had lost macrolide and tetracycline resistance, while harbouring the ins and xis genes of the Tn916–Tn1545 family of conjugative transposons, was documented. Two different tet(M) alleles were detected in isolates of the clone, one of them with a mosaic structure.

Conclusions: The finding of different patterns or alleles of the genes responsible for antibiotic resistance among isolates of the Spain14-5 clone from different Spanish cities indicates different evolutionary events within isolates of a unique Streptococcus pneumoniae clone.

Keywords: Multilocus sequence typing (MLST) , resistance genes , major clones


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