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Prevalence and clonal characterization of Streptococcus pyogenes clinical isolates with reduced fluoroquinolone susceptibility in Spain.

Antimicrobial agents and chemotherapy (2009-10-07)
Milagrosa Montes, Esther Tamayo, Beatriz Orden, Julián Larruskain, Emilio Perez-Trallero
ABSTRACT

The aim of this study was to determine the prevalence and characteristics of non-fluoroquinolone (FQ)-susceptible Streptococcus pyogenes isolates and to study their mechanisms of resistance. We performed a prospective prevalence study with 468 isolates collected from 2005 to 2007 and a retrospective study that was based on the examination of existing data collected from 1999 to 2008. The retrospective study included data for isolates with high-level resistance (HR) to ciprofloxacin (MIC >or= 32 microg/ml) (HR isolates) and isolates with the same emm types as those reported in the literature with low-level resistance (LR) to ciprofloxacin (MICs, 2 to 8 microg/ml) (LR isolates, n = 205). Genetic characterization of the isolates was performed by means of emm typing and multilocus sequence typing. The prevalence of LR ranged from 1.9% in 2005 to 30.8% in 2007. This increase was mainly due to the circulation of an emm6 subtype (emm6.4) that represented 77.1% of the LR isolates in 2007. Notably, another emm6 subtype, also detected in 2007 (emm6.37), showed coresistance to 14- and 15-membered macrolides mediated by the mefA gene. Only three HR isolates were detected (isolates emm68.1/ST247/T3,13,B3264, emm77/ST399/T28, and emm28/ST52/T28), and all were identified in the retrospective study. Overall, the 673 isolates represented 25 emm types. All LR isolates were clustered into two emm types: emm6 (six emm6 subtypes) and emm75. All the 156 emm6 isolates had LR, harbored the Ser79/Ala mutation in the parC gene product, and had the same sequence type (ST), ST382. Most (21/33) of the emm75 isolates had LR, showed the Ser79/Phe plus Asp91/Asn double mutation in the parC gene product, and were ST150. The Asp91/Asn mutation by itself did not confer resistance to FQs.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ofloxacin, fluoroquinolone antibiotic
Supelco
Norfloxacin, analytical standard, ≥98% (TLC)
Sigma-Aldrich
Ciprofloxacin, ≥98% (HPLC)
Supelco
Ciprofloxacin, VETRANAL®, analytical standard
Supelco
Norfloxacin, VETRANAL®, analytical standard
Supelco
Ofloxacin, VETRANAL®, analytical standard
Sigma-Aldrich
Levofloxacin, 98.0-102.0% anhydrous basis (HPLC)
Sigma-Aldrich
Tetracycline, 98.0-102.0% (HPLC)