Emergence of linezolid resistance in the vancomycin-resistant Enterococcus faecium multilocus sequence typing C1 epidemic lineage

A relatively high rate of vancomycin-resistant Enterococcus faecium not susceptible to linezolid was observed in intensive care unit patients. Linezolid-resistant isolates carried the G2576T mutation in the 23S rRNA gene, belonged to different clones, and shared the same allelic profile, which clusters in the C1 multilocus sequence typing epidemic lineage.     

Emergence of VanD-type vancomycin-resistant Enterococcus faecium in Stockholm, Sweden

A vancomycin-resistant Enterococcus faecium isolate from the urine of a liver transplant patient in Stockholm was found to contain a vanD gene. The sequence of the vanD PCR product shared 100% identity with the vanD5 allele. The isolate was resistant to a relatively high level of vancomycin (128 mg/L) and a low level of teicoplanin (4 mg/L). This is the first VanD-type vancomycin-resistant E. faecium isolate reported in Sweden. The emergence of this strain reinforces the necessity of infection control efforts to interrupt the spread of these organisms.     

Emergence of clonal complex 17 Enterococcus faecium in The Netherlands

The global emergence of vancomycin-resistant Enterococcus faecium has been characterized as the clonal spread of clonal complex 17 (CC17) E. faecium. CC17 was defined upon multilocus sequence typing and is characterized by resistance to quinolones and ampicillin and the presence of the enterococcal surface protein (Esp) in the majority of isolates. The recently noticed increased incidence of vancomycin-susceptible CC17 E. faecium infections in our hospital initiated a nationwide study to determine ecological changes among enterococcal infections. The data and strain collections were obtained from 26 (38%) and 9 (14%) of 66 microbiology laboratories in The Netherlands. E. faecium and E. faecalis were distinguished by multiplex PCR; all E. faecium isolates were genotyped by multiple-locus variable-number tandem-repeat analysis (MLVA), and the presence of esp was identified by PCR. Average numbers of ampicillin-resistant enterococcal isolates from normally sterile body sites per hospital increased from 5 ± 1 in 1994 to 25 ± 21 in 2005. Among all enterococcal bloodstream infections, the proportions of ampicillin-resistant E. faecium (AREF) increased from 4% in 1994 to 20% in 2005 (P < 0.001). All E. faecalis isolates were susceptible to ampicillin, whereas 78% of the E. faecium isolates were resistant (49% of these contained esp). Genotyping revealed that 86% of AREF isolates belonged to CC17, including four dominant MLVA types found in ≥3 hospitals, accounting for 64% of the AREF isolates. Infections caused by CC17 E. faecium has increased nationwide, especially in university hospitals due to the clonal spread of four MLVA types, and seems associated with acquisition of the esp gene.     

Molecular detection of linezolid resistance in Enterococcus faecium and Enterococcus faecalis by use of 5' nuclease real-time PCR compared to a modified classical approach

A nucleotide transversion from guanine to uracil in the 23S rRNA confers linezolid resistance. We describe a real-time PCR using two Taqman probes that detects a single mutated allele among the genomes of Enterococcus faecium and Enterococcus faecalis. Results were confirmed by a classical approach involving LabChip technology assayed with an Agilent Bioanalyzer 2100.     

Chloramphenicol treatment for vancomycin-resistant Enterococcus faecium bacteremia

Objective   To evaluate the results of treating vancomycin-resistant Enterococcus faecium (VREF) bacteremia with chloramphenicol.
Methods   We retrospectively reviewed the charts of all adult patients with VREF bacteremia treated with chloramphenicol during the calendar year 1998 at a 522-bed tertiary referral center in New York City. Patients were identified by reviewing microbiology laboratory records. Patients with clinically significant VREF bacteremia who received chloramphenicol for at least 48 h were included in the study. Clinical and microbiological outcomes were determined. Microbiological and molecular tests were performed on a small representative sample of isolates to identify the presence of resistance mechanisms and to look for similarity among the isolates.
Results   Seven episodes of significant VREF bacteremia occurred in six patients. Mean age was 54 years. All patients had cancer and three had severe neutropenia. Five of seven episodes were associated with chronic indwelling devices, but in only one of these cases was the device removed. All isolates were susceptible to chloramphenicol in vitro. All six microbiologically evaluable episodes had a favorable response to chloramphenicol treatment, and four of seven (57%) clinically evaluable episodes had favorable outcomes. Only one death may have been due to VREF bacteremia, so the maximal attributable mortality was 14%. The three representative samples that were tested further were indistinguishable from one another and they displayed no evidence of resistance mechanisms.
Conclusions   In a cohort of severely ill cancer patients, chloramphenicol was effective in treating VREF bacteremia. The use of chloramphenicol should be considered in treating infections with this highly resistant organism, where therapeutic options are limited.     

Antimicrobial resistance classification using MALDI-TOF-MS is not that easy: lessons from vancomycin-resistant Enterococcus faecium

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Nosocomial spread of Enterococcus faecium resistant to vancomycin and linezolid in a tertiary care medical center

In May 2004 our institution encountered its first clinical isolate of linezolid-resistant, vancomycin-resistant Enterococcus faecium (LRVRE). Between October 2004 and July 2005, 40 patients from whom LRVRE organisms were recovered in clinical specimens were characterized. Epidemiologic investigation and pulsed-field gel electrophoresis patterns indicated a clonal outbreak related to nosocomial spread.     

Vancomycin-Variable Enterococcus faecium: In Vivo Emergence of Vancomycin Resistance in a Vancomycin-Susceptible Isolate

We report the emergence of vancomycin resistance in a patient colonized with a vanA-containing, vanRS-negative isolate of Enterococcus faecium which was initially vancomycin susceptible. This is a previously undescribed mechanism of drug resistance with diagnostic and therapeutic implications.     

Enterococcus faecium in hospitals

Most of the characteristics that have ensured the success of enterococci as nosocomial pathogens were described early in this century.Enterococcus faecium andEnterococcus faecalis, the enterococci most frequently isolated from clinical material, differ fundamentally. The intrinsic antimicrobial resistance ofEnterococcus faecium, supplemented by acquired resistance mechanisms, can generate a glycopeptide-multiply-resistant nosocomial pathogen that survives on hands and in the environment, and has the potential for intra-hospital and inter-hospital spread. The use of terms such as an enterococcus, faecal streptococci and group D streptococci have hindered, and still hinder, our understanding of a species rapidly emerging as the most problematic of nosocomial pathogens.     

Mechanisms of resistance to imipenem in imipenem-resistant, ampicillin-sensitive Enterococcus faecium

Enterococcus faecium has six penicillin-binding proteins (PBP), where PBP5 seems to be the main target for beta-lactam antibiotics. The PBP profiles of three imipenem-resistant, ampicillin-sensitive E. faecium strains, isolated from the same patient, were studied using biotinylated ampicillin and chemiluminescence detection. Imipenem resistance in these strains was found to be associated with hyperproduction of PBP5 compared to the ampicillin- and imipenem-susceptible strain ATCC 19434. PBP5 in the imipenem-resistant strains (S1, B2) exhibited a selectively decreased affinity for imipenem. An 854 bp DNA fragment, corresponding to the penicillin-binding domain of pbp5fm, was studied in the resistant strains and the reference strain. Four amino acid substitutions were observed in the resistant strains compared to the susceptible one. The contribution of these substitutions to the increased production of PBP5 in these strains is unclear since the substitution was observed also in a strain without increased production of PBP5. Our results suggest that the moderate imipenem resistance observed in these strains is associated with increased production of PBP5 with relatively decreased affinity for imipenem, and that evolution of imipenem resistance in E. faecium is dinstinct from that of the other beta-lactams such as ampicillin.     

VanB phenotype-vanA genotype Enterococcus faecium with heterogeneous expression of teicoplanin resistance

Six VanB phenotype-vanA genotype isolates of Enterococcus faecium with heterogeneous expression of teicoplanin resistance which gave rise to an outbreak at a Korean tertiary care teaching hospital have IS1216V in the coding region of vanS. This could be the underlying cause of the VanB phenotype-vanA genotype with heterogeneous expression of teicoplanin resistance.     

Incomplete cross resistance against ionophores in Enterococcus faecium and Enterococcus faecalis strains from pigs and poultry

Thirty-two Enterococcus faecium strains and 33 Enterococcus faecalis strains were tested for their susceptibility to the ionophore antibiotics salinomycin, narasin, monensin, and lasalocid. Enterococcal strains originated from poultry in which these products are in use as coccidiostats, and from pigs in which these products are allowed as growth promoters. Resistance against salinomycin and narasin in enterococci was frequent among poultry strains, whereas in pig strains, resistance was less common. No resistance was found against monensin and lasalocid. Full cross resistance between salinomycin and narasin was evident. There was no cross resistance between these two ionophores and monensin and lasalocid.     

Clinical characteristics and treatment outcomes of vancomycin-resistant Enterococcus faecium bacteremia

Background

Vancomycin-resistant Enterococcus faecium (VRE-fm) bacteremia causes significant mortality in hospitalized patients. We sought to investigate clinical characteristics, treatment outcomes, and microbiological eradication associated with VRE-fm bacteremia.

Streptogramin resistance among Enterococcus faecium isolated from production animals in Denmark in 1997

The genetic background for streptogramin resistance was examined in Enterococcus faecium isolated from pigs (n = 55) and broilers (n = 207) in 1997 in Denmark. Fifty-one percent and 67%, respectively, of the isolates were resistant to streptogramins. Among streptogramin-resistant E. faecium (SREF), the genetic background for streptogramin A resistance could be determined in 96% of the isolates from broilers, compared with 14% among SREF from pigs. For broiler isolates 89% of SREF contained the vat(E) gene and 10% the vat(D) gene. Three of these isolates contained both resistance genes. Among SREF from pigs two isolates contained the vat(E) gene and two others the vat(D) gene. The genetic background for streptogramin B was most often identified as the erm(B) gene encoding macrolide, lincosamide, and streptogramin B (MLSB) resistance. Among SREF, 84% and 86% of isolates from broilers and pigs, respectively, contained the erm(B). In SREF from broilers, the erm(B) gene was physically linked to the vat(E) gene in 62% of the vat(E)-positive isolates and 79% of the isolates containing vat(D). erm(A) was detected in two SREF of broiler origin. Both isolates also contained the erm(B) gene. No SREF contained the vgb(A) gene encoding streptogramin B resistance. On the basis of genetic characterization, streptogramin-resistant isolates from broiler were divided into subgroups, according to the presence of the streptogramin A genes, to determine possible co-resistance to antimicrobials, especially glycopeptides. Twenty-five percent of the SREF from broilers were glycopeptide resistant (MIC > 16 microg/ml). None of the isolates containing the streptogramin A gene vat(D) was resistant to glycopeptide, whereas isolates containing the vat(E) gene had a lower prevalence to glycopeptide resistance than the streptogramin-sensitive isolates.     

Detection of the classical G2576U mutation in linezolid resistant Staphylococcus aureus along with isolation of linezolid resistant Enterococcus faecium from a patient on short-term linezolid therapy: First report from India

Purpose: Linezolid is an effective drug against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). We describe the emergence of linezolid resistance in MRSA and VRE from India. Material and Methods: One MRSA and two VRE strains were isolated from a patient on linezolid therapy of one week duration. All three isolates were resistant to linezolid with minimal inhibitory concentrations (MIC) ≥4 mg/L. The 746-bp region flanking the possible G2576U mutation on the corresponding DNA from the 23S rRNA was amplified by polymerase chain reaction (PCR) and amplicons were sequenced for all the three isolates. Conjugation experiments using the linezolid resistant MRSA (LRMRSA) and linezolid resistant VRE (LRVRE) isolates as donors and wild strains of corresponding genera as recipients were performed. Results: The MRSA isolate had the classical G2576U mutation. High quality value scores in the sequencing software validated the mutation. Conjugation studies did not indicate presence of transferable resistance for linezolid. Sequencing did not indicate presence of any mutation in the two LRVRE isolates. Conclusions: This is the first report from India citing resistance in Staphylococcus and Enterococcus against Linezolid.     

Vancomycin-Resistant Enterococcus faecium Colonization in Children

Nosocomial vancomycin-resistant Enterococcus (VRE) infections have been described in only small numbers of pediatric patients. In none of these studies were multivariate analyses performed to assess which factors were independent risk factors in these patients. In the present cohort study of patients admitted to our hematology/oncology unit, surveillance cultures revealed a colonization rate of 24% and all isolates were identified as Enterococcus faecium. Risk factors associated with colonization with VRE identified by multiple logistic regression analysis included young age and chemotherapy with antineoplastic agents, cefotaxime, vancomycin, and ceftazidime. A molecular epidemiological tool, pulsed-field gel electrophoresis, was used to determine the relatedness of the VRE isolates detected. DNA analysis by this method identified two major clusters of VRE isolates. Young children with gastrointestinal colonization with VRE, without evidence of clinical infection, can serve as a reservoir for the spread of VRE.     

Heteroresistance to vancomycin in Enterococcus faecium

This study presents the first report of vancomycin heteroresistance in an Enterococcus faecium isolate from a patient. The original isolate was susceptible in vitro to vancomycin. E-tests showed growth of subcolonies in a zone of inhibition with a vancomycin MIC of >256 microg/ml. Both the susceptible and resistant colonies were from the same strain as determined by PFGE, and both contained the vanA gene as determined by PCR.