Identification of uropathogenic Escherichia coli clonal group A (CgA) in hospitalised patients

Abstract

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...

Diversity and biofilm-production ability among isolates of Escherichia coli phylogroup D belonging to ST69, ST393 and ST405 clonal groups

“…Isolates belonging to cluster I seem to have been circulating among different continents since at least 1999, as reflects this and other studies [31-33]. Despite of the small sample analysed, differences among ST69 isolates from human and non-human origins suggest independent evolution of particular E .…”

Detection of Clonal Group A Escherichia coli Isolates from Broiler Chickens, Broiler Chicken Meat, Community-Dwelling Humans, and Urinary Tract Infection (UTI) Patients and Their Virulence in a Mouse UTI Model

“…Thus, we cannot exclude such isolate origins as possible reservoirs. The detection of CgA isolates from UTI patients also supported previous findings of a widespread global emergence of CgA isolates among uropathogenic E. coli (UPEC) (2,3,(10)(11)(12)(14)(15)(16)(17).…”

“…The possibility that the PFGE profiles of one community-dwelling human and two UTI isolates are related to the PFGE profile of the ATCC BAA-457 isolate is surprising considering the geographical distance and absence of any apparent epidemiological connections. However, other studies have likewise reported PFGE similarities of UTI isolates obtained in Chicago, IL, and Curitiba and Rio de Janeiro, Brazil, to the ATCC BAA-457 isolate (2,3,12). We have no explanation for this finding, but there is a possibility that the BAA-457 isolate belongs to an epidemic, broadly disseminated E. coli lineage.…”

“…Recent studies have suggested that animals and food may be the source of antimicrobial-resistant extraintestinal pathogenic E. coli isolates (19). The distribution of CgA isolates has mainly been investigated in North America, although a few CgA isolates have been identified among UTI E. coli isolates from Europe, South America, and the Middle East (2,3,(10)(11)(12)(14)(15)(16)(17). Furthermore, the sources of CgA isolates have received little investigation.…”

Prevalence and multidrug resistance of Escherichia coli from community-acquired infections in Lagos, Nigeria

“…In the United States, prevalence of MDR E. coli among community isolates increased more than 56% from the 1950s to the 2000s [1]. In Latin America, the widespread dissemination of phylogenetically related strains of uropathogenic E. coli in community and hospital settings has been observed and attributed to a common source in the community such as food and water or person-to-person transmission [16,17].…”