Genotypic characterization of vancomycin-resistant Enterococcus causing urinary tract infection in northern India

Background & objectives: Infections caused by vancomycin-resistant Enterococci are difficult to treat given the limited therapeutic alternatives. Different gene clusters are known to confer vancomycin resistance. vanA and vanB genes are transferable and are clinically relevant. This cross-sectional study aimed to identify the vancomycin-resistant genotypes in the strains causing urinary tract infection and also to test the in vitro efficacy of linezolid and pristinamycin against the vancomycin-resistant isolates. Methods: Antimicrobial resistance profile of 118 enterococcal isolates was evaluated. Minimum inhibitory concentration of vancomycin, teicoplanin and high-level gentamicin (HLG) was determined by micro broth dilution. The vancomycin-resistant isolates were tested against linezolid and pristinamycin by micro-broth dilution and E strip method. The presence of vancomycin-resistant genes was detected by multiplex polymerase chain reaction and was sequenced and analyzed. Results: Most commonly isolated species were Enterococcus faecalis (76.9%) and Enterococcus faecium (16.9%). It was found that 43 per cent of the isolates were resistant to HLG and 16.9 per cent to vancomycin. Higher resistance was seen against fluoroquinolones, erythromycin, tetracycline and ?-lactam drugs. However, 5.08 per cent strains were resistant to tigecycline. All vancomycin-resistant strains were sensitive to pristinamycin and one was resistant to linezolid. vanA and vanB gene were found in 15 and five isolates, respectively. The gene sequences were submitted to NCBI gene bank and accession numbers were obtained. Interpretation & conclusions: The present study showed prevalence of vanA and vanB genes carrying Enterococcus in a tertiary care centre in north India. The emergence of resistance against drugs such as tigecycline and linezolid is a topic of concern as it will be a therapeutic challenge for physicians.

Resistencia de cepas de Staphylococcus aureus aislados en ambientes nosocomiales / Resistance of Staphylococcus aureus strains isolated in nosocomial environments / Resistência de cepas de Staphylococcus aureus isoladas em ambientes nosocomiais

Staphylococcus aureus es un microorganismo que posee características particulares de virulencia y resistencia a los antibióticos. Las infecciones que produce, ocurren mayormente en personas inmunodeprimidas que podrían presentar severas consecuencias, a pesar de la terapia antimicrobiana. Objetivo. Determinar la resistencia de Staphylococcus aureus aislados en ambientes nosocomiales mediante métodos convencionales y moleculares. Materiales y métodos. el estudio presenta un enfoque cuantitativo, investigación de campo, observacional de corte transversal. Se analizó 200 muestras de aislados de ambientes nosocomiales mediante métodos fenotípicos (antibiograma por la técnica de Kirby Bauer) y genotípicos (genes de resistencia blaZ, mecA, y vanA por PCR punto final). Resultados. Los resultados de las pruebas de susceptibilidad antibiótica mostraron que el 100% de las cepas de S. aureus aisladas, fueron resistentes a oxacilina y penicilina g; y sensibles a vancomicina. Conclusiones. los resultados obtenidos en este estudio evidenciaron la presencia de SARM en las diferentes áreas hospitalarias, constituyéndose en un factor de riesgo de transmisión horizontal entre el personal sanitario y los pacientes, razón por la cual es de gran importancia evaluar su prevalencia y establecer medidas rigurosas de prevención y control de su diseminación, para disminuir el riesgo de nuevas infecciones.

Staphylococcus aureus is a microorganism with particular characteristics of virulence and resistance to antibiotics. The infections it produces occur mostly in immunocompromised individuals who may present severe consequences, despite antimicrobial therapy. Objective. To determine the resistance of Staphylococcus aureus isolated in nosocomial environments by conventional and molecular methods. Materials and methods. The study presents a quantitative, field research, observational, cross-sectional approach. Two hundred samples of isolates from nosocomial environments were analyzed by phenotypic (antibiogram by Kirby Bauer technique) and genotypic (resistance genes blaZ, mecA, and vanA by endpoint PCR) methods. Results. The results of the antibiotic susceptibility tests showed that 100% of the S. aureus strains isolated were resistant to oxacillin and penicillin g; and sensitive to vancomycin. Conclusions. the results obtained in this study showed the presence of MRSA in the different hospital areas, constituting a risk factor for horizontal transmission between health personnel and patients, which is why it is of great importance to evaluate its prevalence and establish rigorous measures for the prevention and control of its dissemination, in order to reduce the risk of new infections.

O Staphylococcus aureus é um microorganismo com características particulares de virulência e resistência a antibióticos. As infecções ocorrem principalmente em indivíduos imunocomprometidos que podem ter conseqüências graves, apesar da terapia antimicrobiana. Objetivo. Para determinar a resistência de Staphylococcus aureus isolado em ambientes nosocomiais usando métodos convencionais e moleculares. Materiais e métodos. O estudo apresenta uma abordagem quantitativa, pesquisa de campo, observacional, transversal. Duzentas amostras de isolados de ambientes nosocomiais foram analisadas pelos métodos fenotípico (antibiograma pela técnica de Kirby Bauer) e genotípico (genes de resistência blaZ, mecA, e vanA pelo método de PCR de ponto final). Resultados. Os resultados dos testes de suscetibilidade aos antibióticos mostraram que 100% das cepas de S. aureus isoladas eram resistentes à oxacilina e penicilina g; e sensíveis à vancomicina. Conclusões. Os resultados obtidos neste estudo mostraram a presença de MRSA nas diferentes áreas hospitalares, constituindo um fator de risco para a transmissão horizontal entre o pessoal de saúde e os pacientes, razão pela qual é de grande importância avaliar sua prevalência e estabelecer medidas rigorosas para a prevenção e controle de sua disseminação, a fim de reduzir o risco de novas infecções.

Vigilancia epidemiológica de Staphylococcus aureus y resistencia antibiótica en ambientes nosocomiales / Epidemiological surveillance of Staphylococcus aureus and the antibiotic resistence in nosocomial environments / Vigilância epidemiológica de Staphylococcus aureus e resistência a antibióticos em ambientes nosocomiais

Uno de los microrganismos más importantes en Infecciones Asociadas a la Atención en Salud (IAAS) es Staphylococcus aureus, una bacteria aerobia Gram positiva, resistente a diferentes condiciones ambientales. Objetivo. Identificar Staphylococcus aureus y su resistencia a los principales antibióticos Betalactámicos, aislada en áreas inertes. Materiales y métodos. Se realizó análisis fenotípico, antibiograma y métodos moleculares como: Extracción de ADN mediante Lisis Alcalina, identificación molecular para la amplificación de los genes tanto de identificación de la bacteria (nucA y femB), como de resistencia de antibióticos (blaZ, mecA y vanA) mediante PCR punto final, la separación de los amplicones se realizó mediante electroforesis en gel de Agarosa, los productos de la PCR se revelaron mediante la utilización de transiluminador UV. Resultados. De 200 muestras tomadas se obtuvo dos muestras positivas (1%) para Staphylococcus aureus, con el 100% de resistencia a penicilina y sensible a todos los demás antibióticos testeados. Conclusiones. La identificación de la bacteria y su resistencia hoy en día se realiza mayormente mediante métodos moleculares, lo cual no descarta la identificación fenotípica que, en este caso, determinó resultados importantes como lo es la prueba D-test positivo. La resistencia a fármacos betalactámicos se considera como un serio problema de salud, por lo tanto, se requiere de una vigilancia epidemiológica constante.

Staphylococcus aureus is one of the most important microorganisms related to Health Care Associated Infections (HCAI), it is a Gram-positive aerobic bacterium, highly resistant to the outer environment. Objective. Identify Staphylococcus aureus and its resistance to the most common beta-lactam antibiotics in isolated, inert areas. Materials and methods. Phenotypic analysis, antibiogram and molecular testing such as: DNA extraction by Alkaline Lysis, molecular identification for the amplification of genes both for identification of Staphylococcus aureus (nucA and femB), and for antibiotic resistance (blah, mega and vanA) by PCR, the disassociation of the amplicons was preforming by Agarose gel electrophoresis and results were shown in a UV translluminator. Results. From the 200 samples taken, two showed positive (1%) for Staphylococcus aureus, with 100% penicillin-resistant and sensitive to all other antibiotics tested. Conclusions. Nowadays identification of the bacteria and its resistance is carried out mostly through molecular testing, ruling out phenotypic identification, which in this case it determined important results such as the positive D-test. Resistance to beta-lactam drugs is considered a serious health issue, therefore it requires a safer epidemiological vigilance, an increase in sensitivity testing and the accuracy of results. It is recommended to carry out the D-test in laboratories to identify resistance mechanisms and to guide health personnel to select the best option regarding specific and effective treatment for patients affected with MRSA.

Um dos microrganismos mais importantes em Infecções Associadas à Saúde (HAIs) é o Staphylococcus aureus, uma bactéria aeróbica gram-positiva resistente a diferentes condições ambientais. Objetivo. Identificar Staphylococcus aureus e sua resistência aos principais antibióticos beta-lactam, isolados em áreas inertes. Materiais e métodos. Análise fenotípica, antibiograma e métodos moleculares foram realizados tais como: extração de DNA por Alkaline Lysis, identificação molecular para a amplificação dos genes tanto da identificação da bactéria (nucA e femB), e resistência a antibióticos (blaZ, mecA e vanA) pelo ponto final da PCR, a separação dos amplicons foi realizada por eletrofose em gel de Agarose, os produtos PCR foram revelados usando transiluminador UV. Resultados. De 200 amostras colhidas, duas amostras positivas (1%) foram obtidas para o Staphylococcus aureus, com 100% de resistência à penicilina e sensível a todos os outros antibióticos testados. Conclusões. A identificação da bactéria e sua resistência hoje é realizada principalmente por métodos moleculares, o que não exclui a identificação fenotípica que, neste caso, determinou resultados importantes como o teste D positivo. A resistência aos medicamentos beta-lactam é considerada um grave problema de saúde, portanto, é necessária uma vigilância epidemiológica constante.

Numerical Simulation of Hemodynamics in Three Kinds of Convertible Vena Cava Filters / 医用生物力学

Objective To study the hemodynamic effect of the convertible vena cava filters on treating pulmonary embolism with different thrombus diameters and contents. Methods Three kinds of simulated filter models with the same diameter but different filtering structures (L-style, S-style and W-style) were built and then the hemodynamics of the filter after its implantation into the vessels was analyzed by using computational fluid dynamic (CFD) method. Results Without thrombus in the vessels, three kinds of filters in blood had some obstructive effects and increased the average outlet velocity. While the L-style filter caused the maximum average outlet velocity, the S-style filter was in the middle, and the W-style filter was the lowest. Under the condition of thrombus, the structures of the filter rods had no obvious effect on the average outlet velocity of blood flow and thrombus, and differential pressure of blood flow and thrombus between inlet and outlet. With the increase of thrombus’s diameter and content, the hemodynamic factors showed varying degrees of decreasing tendency. The wall shear stresses (WSS) on three kinds of filter rods caused by blood flow were in normal ranges, and WSS on the lower end of filter supports, the joints of supports and filter rods were below the minimum value, where thrombosis was easy to occur. Conclusions The hemodynamic effect of three kinds of convertible vena cava filters with different filtering structures, different thrombus diameters and contents in vessels were analyzed by using CFD method, which would provide theoretical references for the design and development of novel filters.

Comparative analysis on thrombus filtration efficiency for three kinds of convertible vena cava filters / 医用生物力学

Objective To investigate filtration efficiency of convertible vena cava filters on treating pulmonary embolism under the condition of different thrombus diameters and contents.Methods Three kinds of convertible vena cava filter models with different filtering unit structures (L-style,S-style,W-style) were constructed to numerically simulate hemodynamics based on computational fluid dynamic (CFD) methods,and their filtration efficiency was comparatively analyzed under the condition of different thrombus diameters (5,10,15 mm) and contents (10％,15％,20％).Results With the increasing of thrombus diameter and content,the volume fraction of thrombus distributed on the filter bars increased and the filtration efficiency of the filter became better.When the thrombus diameter was 5 mm,the S-style filter's filtration efficiency was the best as compared with the other two kinds of filters.When the thrombus diameter was 10 mm,the W-style filter showed the best filtration efficiency.When the thrombus diameter was 15 mm,the S-style and W-style filter showed the same filtration efficiency,which was better than the L-style filter.Conclusions The implantation of vena cava filters will cause hemodynamic changes,and its filtration efficiency is not only related to filtering unit structures,but also closely related to the diameter and content of thrombus.These results provide a theoretical reference basis for the design and clinical choice of the novel convertible vena cava filter.

Comparative analysis on thrombus filtration efficiency for three kinds of convertible vena cava filters / 医用生物力学

Objective To investigate filtration efficiency of convertible vena cava filters on treating pulmonary embolism under the condition of different thrombus diameter and content. Methods Three kinds of convertible vena cava filter models with different filtering unit structures (L-style, S-style, W-style) were constructed to numerically simulate hemodynamics based on computational fluid dynamic (CFD) methods, and their filtration efficiency were comparatively analyzed under the condition of different thrombus diameter (5, 10, 15 mm) and content (10%, 15%, 20%). Results With the increasing of thrombus diameter and content, the volume fraction of thrombus distributed on the filter bars increased and filtration efficiency of the filter became better. When the thrombus diameter was 5 mm, the S-style filter’s filtration efficiency was the best as compared with the other two styles of filters. When the thrombus diameter was 10 mm, the W-style filter showed the best filtration efficiency. When the thrombus diameter was 15 mm, the S-style and W-style filter showed the same filtration efficiency, which was better than the L-style filter. Conclusions The implantation of vena cava filters will cause hemodynamic changes, and its filtration efficiency is not only related to filtering unit structures, but also closely related to the diameter and content of thrombus. These results provide a theoretical reference basis for the design and clinical choice of the novel convertible vena cava filter.

Comparative analysis on thrombus filtration efficiency for three kinds of convertible vena cava filters / 医用生物力学

Objective To investigate filtration efficiency of convertible vena cava filters on treating pulmonary embolism under the condition of different thrombus diameters and contents.Methods Three kinds of convertible vena cava filter models with different filtering unit structures (L-style,S-style,W-style) were constructed to numerically simulate hemodynamics based on computational fluid dynamic (CFD) methods,and their filtration efficiency was comparatively analyzed under the condition of different thrombus diameters (5,10,15 mm) and contents (10％,15％,20％).Results With the increasing of thrombus diameter and content,the volume fraction of thrombus distributed on the filter bars increased and the filtration efficiency of the filter became better.When the thrombus diameter was 5 mm,the S-style filter's filtration efficiency was the best as compared with the other two kinds of filters.When the thrombus diameter was 10 mm,the W-style filter showed the best filtration efficiency.When the thrombus diameter was 15 mm,the S-style and W-style filter showed the same filtration efficiency,which was better than the L-style filter.Conclusions The implantation of vena cava filters will cause hemodynamic changes,and its filtration efficiency is not only related to filtering unit structures,but also closely related to the diameter and content of thrombus.These results provide a theoretical reference basis for the design and clinical choice of the novel convertible vena cava filter.

Comparative analysis on thrombus filtration efficiency for three kinds of convertible vena cava filters / 医用生物力学

Objective To investigate filtration efficiency of convertible vena cava filters on treating pulmonary embolism under the condition of different thrombus diameters and contents.Methods Three kinds of convertible vena cava filter models with different filtering unit structures (L-style,S-style,W-style) were constructed to numerically simulate hemodynamics based on computational fluid dynamic (CFD) methods,and their filtration efficiency was comparatively analyzed under the condition of different thrombus diameters (5,10,15 mm) and contents (10％,15％,20％).Results With the increasing of thrombus diameter and content,the volume fraction of thrombus distributed on the filter bars increased and the filtration efficiency of the filter became better.When the thrombus diameter was 5 mm,the S-style filter's filtration efficiency was the best as compared with the other two kinds of filters.When the thrombus diameter was 10 mm,the W-style filter showed the best filtration efficiency.When the thrombus diameter was 15 mm,the S-style and W-style filter showed the same filtration efficiency,which was better than the L-style filter.Conclusions The implantation of vena cava filters will cause hemodynamic changes,and its filtration efficiency is not only related to filtering unit structures,but also closely related to the diameter and content of thrombus.These results provide a theoretical reference basis for the design and clinical choice of the novel convertible vena cava filter.

Emergence of vanA gene among vancomycin-resistant enterococci in a tertiary care hospital of North - East India.

Background & objectives: Vancomycin-resistant enterococci (VRE) have become one of the most challenging nosocomial pathogens with the rapid spread of the multi-drug resistant strain with limited therapeutic options. It is a matter of concern due to its ability to transfer vancomycin resistant gene to other organisms. The present study was undertaken to determine the emergence of vancomycin-resistant enterococci and the vanA gene among the isolates in a tertiary care hospital of North-East India. Methods: A total of 67 consecutive enterococcal isolates from different clinical samples were collected and identified by using the standard methods. Antibiogram was done by disk diffusion method and VRE was screened by the disk diffusion and vancomycin supplement agar dilution method. The minimum inhibitory concentration (MIC) value for vancomycin was determined by E-test. The VRE isolates were analyzed by PCR for vanA gene. Results: A total of 54 (81%) Enterococcus faecalis and 13 (19%) E. faecium were detected among the clinical isolates and 16 (24%) were VRE. The VRE isolates were multidrug resistant and linezolid resistance was also found to be in three. MIC range to vancomycin was 16-32 μg/ml among the VRE. The vanA gene was found in nine of 16 VRE isolates. Interpretation & conclusions: Emergence of VRE and presence of vanA in a tertiary care hospital setting in North-East India indicate toward a need for implementing infection control policies and active surveillance.

Caracterização molecular de Enterococcus spp. resistentes à vancomicina em amostras clínicas, ambientes aquáticos e alimentos / Molecular characterization of vancomycin-resistant Enterococcus spp. in clinical samples, aquatic environments and foods

Enterococos são ubíquos no ambiente e fazem parte da microbiota do trato gastrintestinal de humanos e animais. A importância dessas bactérias tem sido associada com infecções hospitalares e resistência a múltiplas drogas, principalmente à vancomicina. O objetivo do presente estudo foi realizar a caracterização molecular de cepas de Enterococcus spp. resistentes à vancomicina (VRE) isoladas a partir de amostras coletadas de pacientes hospitalizados, água superficial de rios urbanos e carne de frango comercializada no Brasil. A presença do gene vanA foi confirmada em 20 cepas multirresitentes isoladas durante 1997-2011. Dentre os isolados VRE, 12 cepas foram identificadas como E. faecium e oito como E. faecalis. Cepas de E. faecium isoladas de amostras clínicas e águas foram classificadas como clonalmente relacionadas pelo PFGE, com perfil virulência predominante (acm+, esp+). Adicionalmente, enquanto cepas de E. faecium isoladas dos rios pertenceram aos ST203, ST412 e ST478 (previamente caracterizados como endêmicos em hospitais brasileiros), novos STs foram identificados entre as cepas de E. faecalis (ST614, ST615 e ST616) e E. faecium (ST953 e ST954) isoladas de alimentos. Sequências completas do transposon Tn1546 das cepas clínicas VREfm 320/07 (ST478) e ambiental VREfm 11 (ST412) mostraram Tn1546-like element de ~12800 pb, com um ponto de mutação no gene vanA na posição 7.698 (substituição do nucleotídeo T pelo C) e uma no gene vanX na posição 8.234 (G pelo T). Além disso, uma deleção na extremidade esquerda do Tn1546, e as sequências IS1251 e IS1216E na região intergênica vanHS e vanYX, respectivamente, também foram detectados. A este respeito, a IS1216E na região intergênica vanXY constitui um conjunto de genes previamente relatado em cepas clínicas de VREfm no Brasil, denotando uma característica regional. IS1216E tem sido associada com os genes tcrB e aadE que conferem resistência ao cobre e aminoglicosídeos, em E. faecium e Streptococcus agalactiae, respectivamente. Portanto, essa IS pode contribuir para a rápida aquisição de resistência antimicrobiana entre as espécies de cocos Gram-positivos clinicamente importantes. Os tipos de Tn1546 indistiguíveis que foram identificados no atual estudo isolados de humano e ambientes aquáticos sugerem uma comum partilha de um pool de genes de resistência à vancomicina

Enterococci are ubiquitous in the environment and in the intestinal tract of humans and animals. The importance of these bacteria has been associated with nosocomial infection and multiple resistance to antimicrobial agents, mainly vancomycin. The aim of the present study was to perform molecular characterization of vancomycin-resistant Enterococcus spp. strains (VRE) isolated from hospitalized patients, surface water of urban rivers and retail chicken meat in Brazil. The presence of the vanA gene was confirmed in 20 multidrug-resistant strains isolated in 1997-2011. Among these VRE isolates, (n = 12) were identified as E. faecium and (n = 8) as E. faecalis. E. faecium strains isolated from water and clinical samples were classified as clonally related by PFGE, the predominant virulence profile being (acm+, esp+). Additionally, while E. faecium strains isolated from rivers belonging to ST203, ST412 and ST478 (previously characterized as endemic in Brazilian hospitals), new STs were identified among strains of E. faecalis (ST614, ST615 and ST616) and E. faecium (ST953 and ST954) isolated from food. Complete sequences of transposon Tn1546 from VREfm clinical strain 320/07 (ST478) and environmental strain VREfm 11 (ST412) showed a Tn1546-like element of ~12800 bp, with T7698C vanA and G8234T vanX mutations. Moreover, deletion of the Tn1546 left extremity, and the IS1251 and IS1216E sequence inside the vanHS and vanYX intergenic region, respectively, were also detected. In this regard, the IS1216E sequence inside the vanXY intergenic region constitutes a gene array previously reported for Brazilian VREfm clinical strains alone, denoting a regional characteristic. IS1216E has been associated with tcrB and aadE genes, which confer resistance to copper and aminoglycosides, in E. faecium and Streptococcus agalactiae, respectively. Therefore, IS1216E should contribute to rapid acquisition of antimicrobial resistance among species of the clinically important Gram-positive cocci. On the other hand, Tn1546-like elements were identical among clinical and environmental VREfm isolates, suggesting sharing of a common vancomycin resistance gene pool

Detection of both vanA & vanB genes in vanA phenotypes of Enterococci by Taq Man RT-PCR

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.

Evaluation of the iNtRON VRE vanA/vanB Real-Time PCR Assay for Detection of Vancomycin-Resistant Enterococci

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.

Molecular characterization of vanA-containing Enterococcus from migratory birds: song thrush (Turdus philomelos)

Vancomycin-resistant enterococci (VRE) were detected in two faecal samples (1.3%) of song thrush in Portugal. vanA isolates showed high level vancomycin/teicoplanin resistance, as well as resistance to ciprofloxacin, quinupristin-dalfopristin and cloranfenicol. Thrush can be a reservoir of VRE and transmit these resistant bacteria to other animals including humans.

Prevalence and antimicrobial resistance pattern of multidrug-resistant enterococci isolated from clinical specimens.

Purpose: Vancomycin-resistant enterococci (VRE) pose an emerging problem in hospitals worldwide. The present study was undertaken to determine the occurrence, species prevalence, antibacterial resistance, and phenotypic and genetic characteristics of VRE isolated in Riyadh hospitals, KSA. Materials and Methods: Two hundred and six isolates of enterococcal species were obtained from clinical samples. The antibiotic susceptibility of isolates and minimum inhibitory concentration (MIC) tests for vancomycin and teicoplanin were determined. Molecular typing of VRE isolates was carried out by using pulsed field gel electrophoresis (PFGE) and the resistance genotype was determined by polymerase chain reaction (PCR). Results: VRE accounted for 3.9% of the isolates and were detected mostly in urine, wound and blood specimens isolated from ICU, internal medicine and surgical wards. All strains were identified to species level and were found to consist of E. faecalis (69.2%), E. faecium (11.3%), E. avium (2.1%), E. hirae (0.8%), E. casseliflavus (1.3%) and E. gallinarum (1.3%) species. According to the susceptibility data obtained, 8 (3.9%) out of 206 isolates were found to be VRE (MICs > 32 μg/ml). The vanA, vanB and vanC gene fragments of E. faecalis, E. faecium and E. gallinarum were amplified from isolates and were detected. PFGE patterns of the VRE isolates revealed homogenous patterns with dominant clone suggesting that the strains intrinsic resistance is independent. Conclusions: This study shows an emergence of VRE along with increased rate of multidrug-resistant enterococci in the area of the study. Regular surveillance of antimicrobial susceptibilities should be done regularly and the risk factors should be determined.

Vancomycin resistance among methicillin resistant Staphylococcus aureus isolates from intensive care units of tertiary care hospitals in Hyderabad.

Background & objectives: Multidrug resistant methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of nosocomial and community acquired infections and is on the rise. The glycopeptide vancomycin has been proposed as the drug of choice for treating such infections. The present study aimed at identifying the vancomycin resistance both phenotypically and genotypically among the MRSA isolates from two tertiary care hospitals in Hyderabad, south India. Methods: MRSA were isolated and identified from different clinical samples collected from ICUs of tertiary care hospitals in Hyderabad using conventional methods. Antibiogram of the isolates and vancomycin MIC were determined following CLSI guidelines. vanA was amplified by PCR using standard primers. Results: All vancomycin resistant S. aureus (VRSA) isolates were MRSA. The VRSA isolates were positive for vanA gene, except one which was negative. All VRSA had a vancomycin MIC in the range of 16-64 mg/l. Interpretation & conclusions: The increase in vancomycin resistance among MRSA and excessive use of antimicrobial agents have worsened the sensitivity. Larger studies need to be done in various geographical regions of the country to better define the epidemiology, mechanism of vancomycin resistance in S. aureus and its clinical implications.

Rapid detection of Van genes in rectal swabs by real time PCR in Southern Brazil / Rápida detecção de genes Van em swabs retais por PCR real-time na região sul do Brasil

INTRODUCTION: Laboratory-based surveillance is an important component in the control of vancomycin resistant enterococci (VRE). METHODS: The study aimed to evaluate real-time polymerase chain reaction (RT-PCR) (genes vanA-vanB) for VRE detection on 115 swabs from patients included in a surveillance program. RESULTS: Sensitivity of RT-PCR was similar to primary culture (75 percent and 79.5 percent, respectively) when compared to broth enriched culture, whereas specificity was 83.1 percent. CONCLUSIONS: RT-PCR provides same day results, however it showed low sensitivity for VRE detection.

INTRODUÇÃO: Vigilância com base em detecção laboratorial é um componente importante no controle de enterococos resistentes a vancomicina (ERV). MÉTODOS: Avaliamos procedimento da reação em cadeia da polimerase real time (PCR-RT) (genes vanA-vanB) para detecção de ERV em 115 swabs de pacientes incluídos em um programa de vigilância. RESULTADOS: A sensibilidade do RT-PCR foi semelhante a da cultura primária (75 por cento e 79,5 por cento, respectivamente) quando comparada com a cultura em caldo enriquecido, enquanto a especificidade foi de 83,1 por cento. CONCLUSÕES: O RT-PCR fornece resultados no mesmo dia, contudo mostra baixa sensibilidade para a detecção de VRE.

Resistencia a vancomicina en cepas de enterococcus faecium aisladas en un hospital universitario / Vancomycin resistance in enterococcus faecium strains isolated at a university hospital

Las especies que conforman el género Enterococcus, son cocos Gram positivos, que usualmente habitan el tracto intestinal de humanos y animales, pero que también pueden ser aislados de fuentes ambientales. Son capaces de sobrevivir a un amplio rango de condiciones ambientales adversas de estrés, lo que les permite colonizar un amplio rango de nichos, incluidos los ambientes hospitalarios. La patogenicidad nosocomial de los enterococos ha emergido en los últimos años, así como su incrementada resistencia a los antibióticos glicopéptidos. El presente estudio intenta determinar la resistencia a vancomicina de cepas de Enterococcus faecium aisladas en un hospital universitario en el período enero-diciembre del año 2009, así como caracterizar el determinante genético responsable de la resistencia. Para ello se utilizaron los métodos para determinar susceptibilidad a glicopéptidos descritos por el CLSI, adicionalmente se determinó el elemento genético responsable de la resistencia mediante la reacción en cadena de la polimerasa. El 48,81% de las cepas de E. faecium estudiadas fueron resistentes a vancomicina y el PCR demostró la presencia del gen vanA que confiere altos niveles de resistencia a glicopéptidos. Debido al elevado número de aislados resistentes a vancomicina dentro de la institución se sospecha de la presencia de un brote nosocomial producido por este microorganismo; esta cepa epidémica poseedora del gen vanA posiblemente está dispersa en los diferentes servicios de la institución, por lo que se sugiere realizarestudios epidemiológicos para determinar el rol que este microorganismos juega en la producción de infecciones nosocomiales en la institución estudiada

Genus Enterococcus species are gram positive cocci, usually inhabitants of human and animal intestinal tracts, but can also be isolated from their environmental sources. They are able to survive a wide range of adverse environmental stress conditions, allowing them to colonize a wide range of niches, including hospital environments. Nosocomial pathogenicity of enterococci has emerged in recent years, as well as their increased resistance to glycopeptide antibiotics. This study attempts to determine the resistance to vancomycin of Enterococcus faecium strains isolated in a university hospital during the January-December period of 2009, as well as to characterize the genetic determinant responsible for its resistance. Methods to determine susceptibility to glycopeptides described by CLSI were used; additionally, the genetic elements responsible for resistance were identified using the polymerase chain reaction. Of the E. faecium strains studied, 48.81% were resistant to vancomycin, and PCR showed presence of the vanA gene that confers high levels of resistance to glycopeptides. Due to the large number of isolates resistant to vancomycin, the presence of a nosocomial outbreak produced by this microorganism was suspected; this epidemic strain possessing a vanA gene is possibly scattered in different services of the institution; therefore, epidemiological studies should be performed to determine the role this microorganism plays in the production of nosocomial infections in the institution under study

Vancomycin-dependent Enterococcus faecium vanA: characterization of the first case isolated in a university hospital in Brazil

In this study, we report the characterization of a strain of Enterococcus faecium vanA, which grows only in the presence of vancomycin (VDEfm-UEL). The bacterium was isolated from the feces of a female patient who had undergone surgical treatment of Reinke’s edema and was receiving intravenous vancomycin therapy for infection with methicillin/oxacillin-resistant Staphylococcus aureus, a postoperative complication. Antimicrobial dependence was further confirmed by the vancomycin E-test. VDEfm-UEL was also shown to be resistant to ampicillin, ciprofloxacin, chloramphenicol, erythromycin, levofloxacin, penicillin, rifampicin, and teicoplanin. The putative virulence genes efaA, gelE and esp were detected by PCR. The ddl gene from VDEfm-UEL was cloned and sequenced. Vancomycin dependence seems to be associated with the insertion of a nucleotide in that sequence, which results in a frame-shift mutation, introducing a premature stop codon. This is the first report of vancomycin-dependent E. faecium isolation in a university hospital in Brazil.

First report of vancomycin-resistant Enterococcus faecalis in Uberaba, Minas Gerais state

In this study we report the first isolation of VanA-type vancomycin-resistant Enterococcus faecalis strains from two different patients hospitalized in the same intensive care unit at the hospital of Universidade Federal do Triângulo Mineiro (UFTM), Uberaba, Minas Gerais, Brazil.

Linezolid vancomycin resistant Enterococcus isolated from clinical samples in Tehran hospitals.

Background: Vancomycin-resistant enterococci pose an emerging health risk. The limitation in therapeutic options has resulted in the development of new drugs such as quinupristin/ dalfopristin and linezolid. Aim, Setting and Design: This study investigated the species prevalence and antibacterial resistance among enterococci isolated in selected Tehran hospitals. Materials and Methods: Between March 2006 and August 2007, 200 enterococcal isolates from urine, blood, stool and wound were recovered in 2 teaching hospitals of Tehran province. Susceptibility of all isolates was tested against vancomycin, teicoplanin and linezolid antibiotics by disk diffusion and agar dilution method. Results and Conclusion: Seventeen (8.5%), 6 (3%) and 4 (2%) of the isolates were resistant to vancomycin, teicoplanin and linezolid, respectively. Within the vancomycin-resistant isolates, 6 (35.2%), 4 (25%) and 1 (5.88%) showed vanA, vanB and vanC genotype patterns, respectively. Four (23.5%) of VRE isolates were resistant to linezolid with minimum inhibitory concentrations between 16 and 32 µg/mL. Two linezolid vancomycin resistant enterococci were E. faecium.