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1.
Braz. j. infect. dis ; 21(6): 656-659, Nov.-Dec. 2017. graf
Article in English | LILACS | ID: biblio-1039207

ABSTRACT

ABSTRACT Vancomycin-resistant Enterococcus faecium (VREfm) has emerged as an important global nosocomial pathogen, and this trend is associated with the spread of high-risk clones. Here, we determined the genetic and phenotypic features of 93 VREfm isolates that were obtained from patients in 13 hospitals in Vitória, Espírito Santo, Brazil, during 2012-2013. All the isolates were vancomycin-resistant and harbored the vanA gene. Only 6 (6.5%) of the VREfm isolates showed the ability to form biofilm. The 93 isolates analyzed belong to a single pulsed-field gel electrophoresis lineage and presented six subtypes. MLST genotyping showed that all VREfm belonged to ST412 (the high-risk clone, hospital-adapted). The present study describes the dissemination of ST412 clone in the local hospitals. The clonal spread of these ST412 isolates in the area we analyzed as well as other hospitals in southeastern Brazil supports the importance of identifying and controlling the presence of these microorganisms in health care-related services.


Subject(s)
Humans , Cross Infection/microbiology , Gram-Positive Bacterial Infections/microbiology , Enterococcus faecium/genetics , Vancomycin-Resistant Enterococci/genetics , Bacterial Proteins , Brazil , Microbial Sensitivity Tests , Bacterial Typing Techniques , Enterococcus faecium/drug effects , Electrophoresis, Gel, Pulsed-Field , Multilocus Sequence Typing , Vancomycin-Resistant Enterococci/drug effects , Anti-Bacterial Agents/pharmacology
2.
Braz. j. microbiol ; 48(3): 489-492, July-Sept. 2017. tab, graf
Article in English | LILACS | ID: biblio-889140

ABSTRACT

Abstract The aim of this study was to determine the association between Clostridium difficile (C. difficile) and vancomycin-resistant Enterococcus (VRE) and efficacy of screening stools submitted for C. difficile toxin assay for prevalence of VRE. Between April 2012 and February 2014, 158 stool samples submitted for C. difficile toxin to the Marmara University Microbiology Laboratory, were included in the study. Stool samples were analyzed by enzyme immuno assay test; VIDAS (bioMerieux, France) for Toxin A&B. Samples were inoculated on chromID VRE (bioMerieux, France) and incubated 24 h at 37 °C. Manuel tests and API20 STREP (bioMerieux, France) test were used to identify the Enterococci species. After the species identification, vancomycin and teicoplanin MIC's were performed by E test and molecular resistance genes for vanA vs vanB were detected by polymerase chain reaction (PCR). Of the 158 stool samples, 88 were toxin positive. The prevalence of VRE was 17%(n:19) in toxin positives however, 11.4% in toxin negatives(n:70). All VRE isolates were identified as Enterococcus faecium. These results were evaluated according to Fischer's exact chi-square test and p value between VRE colonization and C. difficile toxin positivity was detected 0.047 (p < 0.05). PPV and NPV were 79% and 47% respectively. In our study, the presence of VRE in C. difficile toxin positives is statistically significant compared with toxin negatives (p < 0.05). Screening for VRE is both additional cost and work load for the laboratories. Therefore VRE screening among C. difficile toxin positive samples, will be cost effective for determination of high risk patients in the hospitals especially for developing countries.


Subject(s)
Humans , Bacterial Toxins/analysis , Clostridioides difficile/metabolism , Clostridium Infections/microbiology , Vancomycin Resistance , Feces/microbiology , Vancomycin-Resistant Enterococci/isolation & purification , Bacterial Toxins/metabolism , Vancomycin/pharmacology , Microbial Sensitivity Tests , Clostridioides difficile/isolation & purification , Clostridioides difficile/drug effects , Clostridioides difficile/genetics , Gram-Positive Bacterial Infections/diagnosis , Gram-Positive Bacterial Infections/microbiology , Clostridium Infections/diagnosis , Vancomycin-Resistant Enterococci/classification , Vancomycin-Resistant Enterococci/drug effects , Vancomycin-Resistant Enterococci/genetics , Anti-Bacterial Agents/pharmacology
3.
Braz. j. microbiol ; 46(1): 161-165, 05/2015. tab, graf
Article in English | LILACS | ID: lil-748243

ABSTRACT

Twenty seven isolates of vancomycin resistant Enterococci based on the disk diffusion and E- test have been screened; being found eight (0.3%) clinical isolates of vanA & vanB through Taq Man Real Time PCR assay. This study shows the presence of both vanA & vanB genotypes in vanA phenotypes clinical isolates in the three hospitals in Iran.


Subject(s)
Humans , Bacterial Proteins/genetics , Carbon-Oxygen Ligases/genetics , Vancomycin-Resistant Enterococci/drug effects , Vancomycin-Resistant Enterococci/genetics , Anti-Bacterial Agents/pharmacology , Disk Diffusion Antimicrobial Tests , Iran , Real-Time Polymerase Chain Reaction
4.
Article in English | WPRIM | ID: wpr-34572

ABSTRACT

BACKGROUND: Recently, the iNtRON VRE vanA/vanB real-time PCR (iNtRON; iNtRON Biotechnology, Korea) assay, a multiplex real-time PCR method, was introduced. In this prospective study, we compared the iNtRON assay with the Seeplex VRE ACE detection kit (Seeplex; Seegene, Korea), a conventional multiplex PCR assay. METHODS: A chromogenic agar-based culture, in which pre-selected vancomycin-resistant enterococci (VRE) was grown and subsequently plated on blood agar with vancomycin disks, was regarded as the reference method. A total of 304 consecutive rectal swab specimens were tested for VRE by culture and by iNtRON and Seeplex PCR assays. For the PCR assays, specimens were enriched for 16-24 hr before PCR. RESULTS: VRE were isolated from 44 (14.5%) specimens by chromogenic agar-based culture. The clinical sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the iNtRON assay were 100% (95% confidence interval: 89.8%-100%), 99.2% (96.9%-99.9%), 95.6% (83.6%-99.2%), and 100% (98.2%-100%), respectively, while those of the Seeplex assay were 97.7% (86.2%-99.9%), 99.6% (97.5%-99.9%), 97.7% (86.2%-99.9%), and 99.6% (97.5%-99.9%), respectively. The iNtRON assay had a detection limit of 3,159 copies/microL and 13,702 copies/microL for the vanA and vanB genes, respectively. No cross-reactivity was observed in 11 non-VRE bacterial culture isolates. CONCLUSIONS: The overall performance of the iNtRON assay was comparable to that of a chromogenic agar-based culture method for prompt identification of VRE-colonized patients in hospitals. This assay could be an alternative or supportive method for the effective control of nosocomial VRE infection.


Subject(s)
Bacterial Proteins/genetics , Bacterial Typing Techniques/methods , Carbon-Oxygen Ligases/genetics , DNA, Bacterial/metabolism , Gram-Positive Bacterial Infections/microbiology , Humans , Reagent Kits, Diagnostic , Real-Time Polymerase Chain Reaction , Vancomycin Resistance/genetics , Vancomycin-Resistant Enterococci/genetics
5.
Mem. Inst. Oswaldo Cruz ; 109(6): 712-715, 09/09/2014. graf
Article in English | LILACS | ID: lil-723999

ABSTRACT

The vanC1 gene, which is chromosomally located, confers resistance to vancomycin and serves as a species marker for Enterococcus gallinarum. Enterococcus faecium TJ4031 was isolated from a blood culture and harbours the vanC1gene. Polymerase chain reaction (PCR) assays were performed to detect vanXYc and vanTc genes. Only the vanXYc gene was found in the E. faecium TJ4031 isolate. The minimum inhibitory concentrations of vancomycin and teicoplanin were 2 µg/mL and 1 µg/mL, respectively. Real-time reverse transcription-PCR results revealed that the vanC1and vanXYc genes were not expressed. Pulsed-field gel electrophoresis and southern hybridisation results showed that the vanC1 gene was encoded in the chromosome. E. faecalis isolated from animals has been reported to harbour vanC1gene. However, this study is the first to report the presence of the vanC1gene in E. faecium of human origin. Additionally, our research showed the vanC1gene cannot serve as a species-specific gene of E. gallinarum and that it is able to be transferred between bacteria. Although the resistance marker is not expressed in the strain, our results showed that E. faecium could acquire the vanC1gene from different species.


Subject(s)
Humans , Bacterial Proteins/genetics , Enterococcus faecium/genetics , Genes, Bacterial/genetics , Vancomycin-Resistant Enterococci/genetics , Anti-Bacterial Agents/pharmacology , Blotting, Southern , Bacterial Proteins/blood , Electrophoresis, Gel, Pulsed-Field , Enterococcus faecalis/genetics , Enterococcus faecium/drug effects , Enterococcus/drug effects , Enterococcus/genetics , In Situ Hybridization/methods , Microbial Sensitivity Tests , Multilocus Sequence Typing , Multigene Family/physiology , Polymerase Chain Reaction , Teicoplanin/pharmacology , Vancomycin Resistance/genetics , Vancomycin/pharmacology
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