BACKGROUND: Accurate and early detection of vancomycin-resistant enterococci (VRE) is critical for controlling nosocomial infection. In this study, we evaluated the usefulness of a selective chromogenic agar medium and of multiplex PCR for detection of VRE, and both these techniques were compared with the conventional culture method for VRE detection. METHODS: We performed the following 3 methods for detecting VRE infection in stool specimens: the routine culture method, culturing in selective chromogenic agar medium (chromID VRE, bioMerieux, France), and multiplex PCR using the Seeplex(R) VRE ACE Detection kit (Seegene Inc., Korea) with additional PCR for vanC genes. RESULTS: We isolated 109 VRE strains from 100 stool specimens by the routine culture method. In chromID VRE, all the isolates showed purple colonies, including Enterococcus gallinarum and E. raffinosus, which were later identified using the Vitek card. All VRE isolates were identified by the multiplex PCR method; 100 were vanA-positive E. faecium, 8 were vanA- and vanC-1-positive E. gallinarum, and 1 was vanA-positive E. raffinosus. CONCLUSIONS: For VRE surveillance, culturing the isolates in chromID VRE after broth enrichment appears to be an accurate, rapid, and easy method for routine screening test. Multiplex PCR is relatively expensive and needs skilled techniques for detecting VRE, but it can be an auxiliary tool for rapid detection of genotype during a VRE outbreak.
Agar/chemistry ; Chromogenic Compounds/*chemistry ; Enterococcus/drug effects/genetics/*isolation & purification ; Enterococcus faecium/genetics/isolation & purification ; Feces/microbiology ; Genotype ; Humans ; Phenotype ; Polymerase Chain Reaction/*methods ; Reagent Kits, Diagnostic ; *Vancomycin Resistance

OBJECTIVETo investigate the homology and resistant mechanism of vancomycin-resistant Enterococci (VRE) isolates.

METHODSA total of 9 VRE isolates were collected from 2006 to 2007 at PUMC hospital. The susceptibility of these isolates to 10 different antibiotics including vancomycin was tested by E-test. These strains were processed by brain heart infusion agar screening in the presence of vancomycin (6 microg/ml), and were analyzed for genotypic characteristics using the multiplex PCR. The homology of the isolates was determined by pulsed-field gel electrophoresis (PFGE).

RESULTSAll the 9 VRE isolates were identified as Enterococci faecium. The visual analysis of PFGE patterns revealed 6 different PFGE types. The vanA gene was confirmed by PCR and sequencing in 9 VRE isolates, which were consistent between phenotype and genotype for glycopeptides resistance.

CONCLUSIONSOnly vanA genotype was detected in PUMC hospital. Clonal dissemination, horizontal gene transfer, and the selective pressure of antimicrobial agents may contribute to the increase of VRE.

Bacterial Proteins ; genetics ; Bacterial Typing Techniques ; Drug Resistance, Multiple, Bacterial ; Enterococcus faecium ; classification ; drug effects ; genetics ; isolation & purification ; Gram-Positive Bacterial Infections ; microbiology ; Humans ; Vancomycin Resistance

INTRODUCTIONUntil recently, vancomycin-resistant enterococcus (VRE) infection or colonisation was a rare occurrence in Singapore. The first major VRE outbreak involving a 1500-bed tertiary care institution in March 2005 presented major challenges in infection control and came at high costs. This study evaluates the predictors of VRE carriage based on patients' clinical and demographic profiles.

MATERIALS AND METHODSStudy patients were selected from the hospital inpatient census population during the VRE outbreak (aged 16 years or more). Clinical information from 84 cases and 377 controls were analysed.

RESULTSSignificant predictors of VRE carriage included: age>65 years Odds ratio (OR), 1.98; 95% CI (confidence interval), 1.14 to 3.43); female gender (OR, 2.15; 95% CI, 1.27 to 3.65); history of diabetes mellitus (OR, 1.94; 95% CI, 1.14 to 3.30), and staying in a crowded communal ward (OR, 2.75; 95% CI, 1.60 to 4.74). Each additional day of recent hospital stay also posed increased risk (OR, 1.03; 95% CI, 1.01 to 1.04).

CONCLUSIONElderly diabetic females with prolonged hospitalisation in crowded communal wards formed the profile that significantly predicted VRE carriage in this major hospital-wide outbreak of VRE in Singapore. It is imperative that active VRE surveillance and appropriate infection control measures be maintained in these wards to prevent future VRE outbreaks.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Cross Infection ; drug therapy ; epidemiology ; microbiology ; Disease Outbreaks ; Enterococcus ; drug effects ; Enterococcus faecalis ; isolation & purification ; Enterococcus faecium ; isolation & purification ; Female ; Humans ; Infection Control ; Male ; Medical Audit ; Middle Aged ; Risk Factors ; Singapore ; epidemiology ; Streptococcal Infections ; drug therapy ; epidemiology ; Vancomycin ; pharmacology ; therapeutic use ; Vancomycin Resistance

BACKGROUND: Few studies have evaluated the performance of the recently introduced MicroScan Synergies plus Positive Combo 3 Panels (SIPC3) (Dade Behring Inc., USA). We evaluated the clinical efficacy of the panels in identification (ID) and antimicrobial susceptibility testing (AST) of Staphylococcusaureus and enterococci. METHODS: To evaluate the panels' accuracy of identification, the results obtained using the test panels were compared with those obtained by using conventional biochemical tests in conjunction with VITEK 2 system (bio-Merieux, USA). In addition, the AST results obtained using the panels were compared with those obtained by performing CLSI broth microdilution. RESULTS: The overall agreement between the approaches for the ID of S. aureus and enterococci was 100% and 96%, respectively. The categorical and essential agreements (CA and EA) for S. aureus were 98%, each. Very major errors (VME), major errors (ME), and minor error (mE) for S. aureus were 0.45%, 0.3%, and 4.2%, respectively. The majority of VMEs were for oxacillin (8.6%), penicillin (2.0%), erythromycin (7.9%), clindamycin (3.8%), and tetracycline (4.1%). For enterococci, the CA, EA, VME, ME, and mE were 88.8%, 93.7%, 4.4%, 0%, and 2.8%, respectively. The 80.5% (29/36) of Enterococcus faecium had concordant ID with the reference. Most of the categorical errors (3 VMEs and 14 mEs) were observed for quinupristin/dalfopristin (Synercid; Catalytica Pharmaceuticals Inc., USA). CONCLUSIONS: The panels compared favorably with conventional methods for the ID and AST of S. aureus. However, we expected a better performance for ID of E. faecium and AST using Synercid.
Anti-Bacterial Agents/*pharmacology ; Clindamycin/pharmacology ; Drug Resistance, Bacterial ; Enterococcus/*drug effects/isolation & purification ; Erythromycin/pharmacology ; Microbial Sensitivity Tests/instrumentation/*methods ; Oxacillin/pharmacology ; Penicillins/pharmacology ; Reagent Kits, Diagnostic ; Staphylococcus aureus/*drug effects/isolation & purification ; Tetracycline/pharmacology

BACKGROUND: To assess the possibility of VRE transmission from animals to humans, we studied the prevalence of vancomycin-resistant enterococci (VRE) in farm animals, raw chicken meat, and healthy people. We then determined the molecular relatedness of VRE isolates between animals and humans in Korea. METHODS: We aimed to isolate VRE from 150 enterococci specimens of farm animals, 15 raw chicken meat samples, and stools from 200 healthy people. Species differentiation was done with conventional biochemical tests. Vancomycin resistance genotyping was done by polymerase chain reaction (PCR). Using the agar dilution method, antimicrobial susceptibility was tested for 8 antimicrobials and pulsed-field gel electrophoresis (PFGE) was done to evaluate the molecular relatedness of VRE isolates. RESULTS: The prevalence of VRE was 14.7% (22/150) in farm animal specimens, 1% (2/200) in healthy people, and 60% (9/15) in raw chicken meat. Of 22 animal VRE isolates, 1 vanA E. faecium, 15 vanC1 E. gallinarum, and 6 vanC2 E. casseliflavus were identified. All of the 9 VRE from raw chicken meat and all of the 20 clinical VRE strains were vanA E. faecium. However, in healthy people, only 2 vanC2 E. casseliflavus were isolated. These showed low-level resistance to vancomycin and susceptibility to teicoplanin. However, 9 VRE strains from raw chicken meat had high-level resistance to vancomycin (MIC50, 90: > 128 microgram/mL), teicoplanin (MIC50, 90: > 128 microgram/mL), ampicillin (MIC50, 90: > 128 microgram/mL), erythromycin (MIC50, 90: > 128 microgram/mL), and tetracycline (MIC50/90: 128/> 128 microgram/mL). CONCLUSION: This study demonstrated little evidence of VRE colonization in healthy people despite high recovery of VRE among raw chicken meat. It is suggested that there is little evidence of VRE transmission from animals to healthy people. However, we assumed that there exists the possibility of VRE contamination during the processing of chicken meat.
Animals ; Cattle/microbiology ; Chickens ; Enterococcus/*drug effects/*isolation & purification ; Feces/microbiology ; Humans ; Korea ; Meat/microbiology ; Prevalence ; Research Support, Non-U.S. Gov't ; Swine/microbiology ; *Vancomycin Resistance

The crude extracts of the fermentation broth from a marine sediment-derived actinomycete strain, Saccharothrix sp. 10-10, showed significant antibacterial activities against drug-resistant pathogens. A genome-mining PCR-based experiment targeting the genes encoding key enzymes involved in the biosynthesis of secondary metabolites indicated that the strain 10-10 showed the potential to produce tetracenomycin-like compounds. Further chemical investigation of the cultures of this strain led to the identification of two antibiotics, including a tetracenomycin (Tcm) analogs, Tcm X (1), and a tomaymycin derivative, oxotomaymycin (2). Their structures were identified by spectroscopic data analysis, including UV, 1D-NMR, 2D-NMR and MS spectra. Tcm X (1) showed moderate antibacterial activities against a number of drug-resistant pathogens, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE) pathogens, with the MIC values in the range of 32-64 microg x mL(-1). In addition, 1 also displayed significant cytotoxic activities against human cancer cell lines, including HL60 (leukemia), HepG2 (liver), and MCF-7 (breast) with the IC 50 values of 5.1, 9.7 and 18.0 micromol x L(-1), respectively. Guided by the PCR-based gene sequence analysis, Tcm X (1) and oxotomaymycin (2) were identified from the genus of Saccharothrix and their 13C NMR data were correctly assigned on the basis of 2D NMR spectroscopic data analysis for the first time.
Actinomycetales ; chemistry ; genetics ; Anti-Bacterial Agents ; chemistry ; isolation & purification ; pharmacology ; Antineoplastic Agents ; chemistry ; isolation & purification ; pharmacology ; Benzodiazepinones ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Data Mining ; methods ; Drug Resistance, Bacterial ; Enterococcus faecalis ; drug effects ; Fermentation ; Genomics ; Humans ; Inhibitory Concentration 50 ; Marine Biology ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; Microbial Sensitivity Tests ; Molecular Structure ; Naphthacenes ; chemistry ; isolation & purification ; pharmacology ; Phylogeny ; Staphylococcus epidermidis ; drug effects

PURPOSE: This study was aimed to describe a vancomycin-resistant enterococci (VRE) outbreak across three intensive care units (ICUs) of a Korean hospital from September 2006 to January 2007 and the subsequent control strategies. MATERIALS AND METHODS: We simultaneously implemented multifaceted interventions to control the outbreak, including establishing a VRE cohort ward, active rectal surveillance cultures, daily extensive cleaning of environmental surfaces and environmental cultures, antibiotic restriction, and education of hospital staff. We measured weekly VRE prevalence and rectal acquisition rates and characterized the VRE isolates by polymerase chain reaction (PCR) of the vanA gene and Sma1-pulsed-field gel electrophoresis (PFGE). RESULTS: During the outbreak, a total of 50 patients infected with VRE were identified by clinical and surveillance cultures, and 46 had vancomycin-resistant Enterococcus faecium (VREF). PFGE analysis of VREF isolates from initial two months disclosed 6 types and clusters of two major types. The outbreak was terminated 5 months after implementation of the interventions: The weekly prevalence rate decreased from 9.1/100 patients-day in September 2006 to 0.6/100 by the end of January 2007, and the rectal acquisition rates also dropped from 6.9/100 to 0/100 patients-day. CONCLUSION: Our study suggests that an aggressive multifaceted control strategy is a rapid, effective approach for controlling a VRE outbreak.
Decontamination ; Disease Outbreaks/*prevention & control ; Enterococcus faecium/*drug effects/isolation & purification ; Gram-Positive Bacterial Infections/drug therapy/*epidemiology/prevention & control ; Humans ; Intensive Care Units ; Patient Isolation ; Prevalence ; *Vancomycin Resistance

INTRODUCTIONVancomycin-resistant enterococci (VRE) have emerged as one of the major nosocomial antimicrobial-resistant pathogens globally. In this article, we describe the epidemiology of VRE in Singaporean public hospitals in the 5 years following the major local VRE outbreak in 2005.

MATERIALS AND METHODSA passive laboratory surveillance programme identified non-duplicate VRE isolates from 7 hospitals from 2006 to 2010. Descriptive statistics and time-series analysis was performed on all clinical VRE isolates for each individual hospital as well as for the combined dataset.

RESULTSThere were a total of 418 VRE isolates over 5 years, of which 102 isolates (24.4%) were from clinical cultures. Between 0.4% and 0.7% of all clinical enterococcal isolates were resistant to vancomycin. The overall incidence-density of VRE did not change over time in Singapore despite 2 separate outbreaks in tertiary hospitals in 2009 and 2010. Incidence-density of clinical VRE cases fell in 2 secondary hospitals, while another 2 hospitals experienced no significant VRE infections after 2008.

CONCLUSIONThe prevalence of VRE clinical isolates remains low in Singaporean public sector hospitals. However, the presence of at least 2 outbreaks in separate hospitals over the past 5 years indicates the need for continued vigilance in order to prevent any further increase in VRE prevalence locally.

Anti-Bacterial Agents ; pharmacology ; Cross Infection ; epidemiology ; Enterococcus ; drug effects ; isolation & purification ; Gram-Positive Bacterial Infections ; drug therapy ; Hospitals, Public ; Humans ; Population Surveillance ; Singapore ; epidemiology ; Vancomycin ; therapeutic use ; Vancomycin Resistance ; drug effects

Monitoring temporal trends of antimicrobial resistance can provide useful information for the empirical selection of antimicrobial agents to treat infected patients and for the control of nosocomial infections. In this study, we analyzed antimicrobial resistance of clinically relevant bacteria in 2003 at Korean hospitals and at a commercial laboratory. The following organism-antimicrobial agent resistance combinations were very prevalent: oxacillin-resistant Staphylococcus aureus (68%), expanded-spectrum cephalosporin-resistant Klebsiella pneumoniae (25%), and fluoroquinolone-resistant Escherichia coli (33%), Acinetobacter spp. (58%), and Pseudomonas aeruginosa (40%). Moreover, gradual increases in vancomycin-resistant Enterococcus faecium (20%), cefoxitin-resistant E. coli (10%) and K. pneumoniae (23%), and imipenem-resistant P. aeruginosa (20%) and Acinetobacter spp. (13%) were also observed. The resistance rates of Acinetobacter spp. to most antimicrobial agents at hospitals and at the commercial laboratory were similar. Among the Acinetobacter spp. isolated at a tertiary-care hospital, 46.2% were multidrug-resistant to 9-12 of 13 antimicrobial agents, and 18.3% were panresistant. The exclusion of duplicate isolates at a tertiary-care hospital significantly lowered the proportion of oxacillin-resistant S. aureus, vancomycin-resistant E. faecium, and fluoroquinolone-resistant E. coli.
Vancomycin Resistance ; Vancomycin/pharmacology ; Population Surveillance ; Microbial Sensitivity Tests ; Korea/epidemiology ; Klebsiella pneumoniae/drug effects/isolation & purification ; Klebsiella Infections/drug therapy/epidemiology/microbiology ; Imipenem/pharmacology ; Humans ; Gram-Positive Bacterial Infections/drug therapy/epidemiology/*microbiology ; Gram-Negative Bacterial Infections/drug therapy/epidemiology/*microbiology ; Gammaproteobacteria/*drug effects/isolation & purification ; Fluoroquinolones/pharmacology ; Enterococcus faecium/*drug effects/isolation & purification ; *Drug Resistance, Bacterial ; Anti-Bacterial Agents/*pharmacology ; Amikacin/pharmacology ; Acinetobacter Infections/drug therapy/epidemiology/microbiology ; Acinetobacter/drug effects/isolation & purification

BACKGROUNDVancomycin-resistant enterococci (VRE) are a major and emerging hospital-acquired pathogen associated with high mortality, particularly among the critically ill and Intensive Care Units (ICUs) patients. This study aimed to determine the prevalence and demographic and clinical characteristics of VRE among patients admitted to a university hospital in Riyadh, Saudi Arabia.

METHODSA study was conducted during the period from September 2014 to November 2015 at King Khalid University Hospital, a tertiary care hospital in Riyadh, Saudi Arabia, including in-patients with VRE infection. Data were collected using laboratory results and the medical records of admitted patients and were analyzed using SPSS version 19.0 statistical software.

RESULTSIn a one-year period, 231 enterococci were isolated from blood, urine, exudates, sputum, stool, and body fluid. There were 191 (82.7%) vancomycin-sensitive enterococci (VSE) and 40 (17.3%) isolates were VRE. The Enterococcus species included E. faecalis 168 (72.7%), E. faecium, 53 (22.8%) E. gallinarum 5 (2.2%), and E. avium 5 (2.2%). VRE were more significant from blood specimens (P< 0.0001) while VSE were significantly more predominant from urine specimens (P< 0.0001). VRE were more commonly isolated from patients in ICUs and oncology unit (P = 0.0151 and P< 0.001, respectively) while VSE were more predominant in the medical and surgical areas (P = 0.0178 and P = 0.0178, respectively).

CONCLUSIONSThis study highlights the high prevalence of VRE in the hospital and the association of enterococcal infections with high-risk areas and oncology units, which warrant more studies looking for better management of these infections.

Adult ; Aged ; Anti-Bacterial Agents ; pharmacology ; Child, Preschool ; Enterococcus ; drug effects ; Female ; Hospitals, Teaching ; Humans ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Saudi Arabia ; Vancomycin ; pharmacology ; Vancomycin-Resistant Enterococci ; drug effects ; isolation & purification ; pathogenicity