Recent studies from North America and Europe have demonstrated community-wide clonal spread of uropathogenic Escherichia coli (UPEC). To investigate if a similar pattern of spread occurs in Brazil, we characterized UPEC from women with community-acquired urinary tract infection (UTI) in Rio de Janeiro. E. coli isolates from women with UTI in one public outpatient clinic were evaluated for antibiotic susceptibility, E. coli phylogenetic grouping, enterobacterial repetitive intergenic consensus (ERIC) 2 PCR and pulsed-field gel electrophoresis fingerprinting, and multilocus sequence typing. From March 2005 to November 2006, 344 patients were studied. Of these, 186 (54%) had confirmed UTI, 118 (63.4%) of which were caused by E. coli. More than 50% of these isolates were resistant to ampicillin and trimethoprim/sulfamethoxazole. Of these, 96 (81%) belonged to 19 ERIC2 clonal groups. The largest group included 15 isolates, all belonging to multilocus sequence typing group ST69 and phylogenetic group D; they had pulsed-field gel electrophoresis patterns sharing at least 89% similarity compared with the CgA reference strain ATCC BAA-457. CgA strains have been found to be widespread in the United States in the early 2000s. Clonal group E. coli strains accounted for a large proportion (52%) of all UTIs and 82% of the trimethoprim/sulfamethoxazole-resistant E. coli UTIs. Thus, as in North America and Europe, UPECs that cause UTI in Rio de Janeiro also show clonal distribution, and a substantial proportion of drug-resistant UTI is caused by a small set of genetically related E. coli strains.
Mupirocin is a topical antimicrobial agent that has been sucessfully used to eradicate methicillin-resistant Staphylococcus aureus from the anterior nares and other sites of patients and health care personnel. This report describes the acquisition of a novel mupirocin resistance gene (iles) by an epidemic MRSA clone that is geographically widespread in Brazil.
This study provides the first description of healthcare-associated infections with (Manges et al. 2001, 2004, Johnson et al. 2002, Petrof et al. 2002. CgA isolation has also been described in other parts of the world (Johnson et al. 2005, Manges et al. 2008. The isolates in Latin America studied to date were found among a collection of 60 E. coli isolates of unknown origin in Curitiba, state of Paraná, Brazil. CgA was found in three urine isolates by random amplification of polymorphic DNA and phylogenetic grouping analyses (Johnson et al. 2005).In the present report, we describe the identification of three CgA E. coli isolates from a public university-affiliated hospital in Rio de Janeiro, Brazil, exhibiting the four predominant ERIC-PCR bands of approximately 1145, 1029, 908 and 720 bp that are typically observed for CgA (Manges et al. 2001). These isolates were further characterised as described below.Initially, E. coli isolates were evaluated by ERIC-PCR and pulsed-field gel electrophoresis (PFGE), as previously described (Bender et al. 1997, Dias et al. 2008. Briefly, for ERIC-PCR, amplifications were carried out in a total volume of 25 μL. Each reaction contained buffer, 0.1 mM of each dNTP, 3 mM MgCl 2 , 0.3 μM ERIC2 primer (5'-AAGTAAGTGACTGGGGTGAGCG-3'), 1.5 U Taq DNA polymerase (BIOTOOLS) and 3 μL template DNA. Reaction conditions were as follows: a 2 min initial denaturation at 94°C followed by 40 cycles of a 30 sec denaturation at 94°C, a 1 min annealing at 54°C and a 4 min extension at 72°C and a final extension step of 1 min at 72°C. Amplification products were electrophoresed on 1.5% agarose gels. For PFGE, chromosomal DNA in the plugs was digested with XbaI (New England BioLabs) at 37°C according to the instructions of the manufacturer. The restriction fragments were separated by PFGE on 1% agarose gels and electrophoresis was carried out in a CHEF DR II system (Bio-Rad Laboratories) at 13°C and 6 V/cm for 22 h with pulse times ranging from 2.2-54.2 sec. DNA-banding patterns were interpreted by visual inspection and with the GelCompar II program version 4.01 (Applied Maths, Sint-Martens-Latem, Belgium). The CgA reference strain ATCC BAA-457 was included for comparison in all experiments.For phylogenetic analysis, E. coli phylogrouping (A, B1, B2 or D) was assessed using a previously reported triplex PCR-based assay (Clermont et al. 2000). In addition, multilocus sequence typing (MLST) was performed using a standardised protocol for E. coli maintained at the MLST Databases at the ERI, University College Cork website (http://mlst.ucc.ie/). Briefly, amplifications were carried out in a total volume of 50 μL. Each reaction contained buffer, 0.2 mM of each dNTP, 1.5 mM MgCl 2 , 0.2 μM of each primer, 2.5 U AmpliTaq Gold (Applied Biosystems) and 2 μL template DNA. Reaction conditions were as follows: a 2 min initial denaturation at 95°C followed by 30 cycles of a 1 min denaturation at 95°C, a 1 min annealing at the temperature specified for each gene (http://mlst.ucc.ie/) and a 2 min extension at 72°C a...
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