Molecular epidemiology and antimicrobial susceptibility of Enterococci recovered from Brazilian intensive care units

Titze‐de‐Almeida, Ricardo; Filho, Maurício Rollo; Nogueira, Celeste A.; Rodrigues, Isabela Pereira; Filho, João Eudes; Nascimento, Rejane Soares do; Ferreira, Renato Fonseca; Moraes, Lídia Maria Pepe de; Boelens, Hélène; Belkum, Alex van; Felipe, Maria Sueli Soares

doi:10.1590/s1413-86702004000300002

Cited by 16 publications

(17 citation statements)

References 36 publications

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“…It also reduced the time usually spent for DNA extraction because the colonies are picked up directly from the plate to the PCR mixture. The rate of agreement between this PCR scheme and the conventional biochemical assays was high for all species tested, eg., E. faecalis (95%), E. faecium (95%), E. gallinarum (100%) and E. casseliflavus (100%) [13]. In the present study, only 4 isolates (9.8%) showed discordance between PCR and the biochemical testing.…”

Section: Activities Timecontrasting

confidence: 45%

“…This method also detects, in the same reaction, the resistance genes vanA and vanB ( Table 1). The primer concentrations previously adjusted to provide easily distinguished bands were ddl E. faecalis , ddl E. faecium , E. gallinarumvanC1, E. casseliflavus-vanC2/3 (18 pmol of each primer), vanA (3 pmol of each primer), and vanB (2 pmol of each primer) [13]. For each PCR reaction, 5 -10 colonies were picked up from the chromogenic medium II.…”

Section: Multiplex Polymerase Chain Reaction Schemementioning

confidence: 99%

“…The method used in our study allows the simultaneous identification of E. faecium, E. faecalis, E. casseliflavus, and E. gallinarum sequences, as well as the vancomycin-resistant genes vanA and vanB in one unique reaction [13]. The scheme used primers previously described and provided easily visualized bands after agarose gel electrophoresis [21,24].…”

Section: Activities Timementioning

confidence: 99%

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Rapid detection of Vancomycin-Resistant Enterococci (VRE) in rectal samples from patients admitted to intensive care units

et al. 2009

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The reduction in time required to identify vancomycin-resistant enterococci (VRE) has gained increased importance during hospital outbreaks. In the present study, we implemented a laboratory protocol to speed up the VRE screening from rectal samples. The protocol combines a medium for selective VRE isolation (VREBAC ® , Probac, São Paulo) and a multiplex PCR for detection and identification of vanA and vanB resistance genes. The screening performance was analyzed in 114 specimens collected from four intensive care units. The swabs were collected at two periods: (1) during a VRE outbreak (February 2006, n=83 patients) and (2) at the post-outbreak period, after adoption of infection control measures (June 2006, n=31 patients). Forty-one/83 VRE (49.4%) and 3/31(9.7%) VRE were found at the first and second period, respectively. All isolates harbored the vanA gene. In both periods, detection of the gene vanA parallels to the minimum inhibitory concentration values of >256 µ µ µ µ µg/mL and >48 µ µ µ µ µg/mL for vancomycin and teicoplanin, respectively. Multiplex PCR and conventional methods agreed in 90.2% for enterococci identification. Besides this accuracy, we also found a remarkable reduction in time to obtain results. Detection of enterococcal species and identification of vancomycin resistance genes were ready in 29.5 hours, in comparison to 72 hours needed by the conventional methods. In conclusion, our protocol identified properly and rapidly enterococci species and vancomycin-resistance genes. The results strongly encourage its adoption by microbiology laboratories for VRE screenning in rectal samples.

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Section: Activities Timecontrasting

confidence: 45%

Section: Multiplex Polymerase Chain Reaction Schemementioning

confidence: 99%

Section: Activities Timementioning

confidence: 99%

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Rapid detection of Vancomycin-Resistant Enterococci (VRE) in rectal samples from patients admitted to intensive care units

et al. 2009

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“…In Brazil, clinical and epidemiological studies of enterococcal infections have been less comprehensive, with most studies focusing on vancomycin-resistant En-High-level gentamicin-resistant E. faecalis bacteremia www.bjournal.com.br terococcus (VRE), which is still restricted to a few Brazilian hospitals (6)(7)(8).…”

Section: Introductionmentioning

confidence: 99%

“…Since Enterococcus spp is the second most common cause of Gram-positive blood stream infections in Brazil and that approximately 76% of all enterococcal infections are due to Enterococcus faecalis, we evaluated cases of bacteremia caused by this organism at the teaching hospital of the Universidade Estadual de Campinas (Unicamp), Campinas, São Paulo State (8). Our objective was to characterize the epidemiological, clinical, and microbiological features of bacteremias caused by HLGR and non-HLGR E. faecalis isolated from January 1999 through December 2003.…”

Section: Introductionmentioning

confidence: 99%

Clinical, epidemiological, and microbiological characteristics of bacteremia caused by high-level gentamicin-resistant Enterococcus faecalis

Vigani

Oliveira

Bratfich

et al. 2008

Braz J Med Biol Res

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Enterococcus spp bacteremia is associated with high mortality and the appearance of high-level gentamicin resistance (HLGR) created additional challenges for the treatment of these infections. We evaluated the epidemiological and clinical characteristics of patients with bacteremias caused by HLGR and non-HLGR Enterococcus faecalis isolates at a teaching hospital in the State of São Paulo, Brazil. Patients with bacteremia due to E. faecalis diagnosed between January 1999 and December 2003 were included in the study. We collected clinical, epidemiological, and microbiological data from medical records. Banked isolates were typed using pulsed-field gel electrophoresis. We identified 145 cases of E. faecalis bacteremia: 66 (45.5%) were caused by HLGR isolates and 79 (54.5%) by non-HLGR. In the univariate analysis, patients with HLGR infection were older, had higher rates of bladder catheterization, and more often had treatment with cephalosporin, quinolone, and/or carbapenem compared with patients with non-HLGR infection (P < 0.05). Multivariate analysis indicated that older age, hematological malignancy, and previous use of vancomycin were independently associated with HLGR (P < 0.05). Mortality rates were not significantly different among patients with HLGR (50%) and non-HLGR (43%) infections (P = 0.40). Of the 32 genotyped isolates, 16 were distributed into 6 main electrophoresis patterns and 16 others had distinct patterns. E. faecalis bacteremia is associated with high mortality and is frequently caused by HLGR isolates at this teaching hospital. The variability among genotyped isolates suggests that endogenous infections, rather than patient-to-patient transmission of E. faecalis, are more common at this institution.

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Virulence determinants in vancomycin-resistant Enterococcus faecium vanA isolated from different sources at University Hospital of Londrina, Paraná, Brazil

et al. 2010

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Enterococcus faecium, especially those showing multidrug resistance, has emerged as a significant cause of healthcare-associated infections worldwide. However, relatively little is known about the virulence and pathogenesis of this species. The aim of this study was to determine the occurrence of four putative virulence determinants of E. faecium and to correlate them with phenotypic traits. Using forty E. faecium vanA-type isolates from hospitalized patients and their environmental vicinity, we determined the following: the antimicrobial susceptibility profile, occurrence of the genes cylA, efaA, esp, and gelE, hemolytic and gelatinase activities, capacity to form biofilm and in vitro adhesion to epithelial cells. All isolates were shown to be resistant to vancomycin and teicoplanin, as well as to two or more other antimicrobials. All isolates harbored at least one putative virulence marker, and the prevalence was as follows: esp, 87.5%; efaA, 82.5%; gelE, 70%; and cylA, 65%. The presence of 4 genes was observed in 32.5% isolates. The presence of the efaA was associated with the presence of esp, regardless of the source of the isolates. A positive association with the presence of cylA and hemolytic activity in the sheep blood agar assay was observed. No association was found for gelE and gelatinase production in the agar plate assay, for efaA and LLC-MK2 cell adhesion, and for esp and biofilm formation on polystyrene surface. These results show the presence of putative virulence genes in multiple antimicrobial resistant E. faecium isolates from different sources in a hospital setting.

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Molecular epidemiology and antimicrobial susceptibility of Enterococci recovered from Brazilian intensive care units

Cited by 16 publications

References 36 publications

Rapid detection of Vancomycin-Resistant Enterococci (VRE) in rectal samples from patients admitted to intensive care units

Rapid detection of Vancomycin-Resistant Enterococci (VRE) in rectal samples from patients admitted to intensive care units

Clinical, epidemiological, and microbiological characteristics of bacteremia caused by high-level gentamicin-resistant Enterococcus faecalis

Virulence determinants in vancomycin-resistant Enterococcus faecium vanA isolated from different sources at University Hospital of Londrina, Paraná, Brazil

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