Characterization of a vancomycin-resistant Enterococcus faecium isolate and a vancomycin-susceptible E. faecium isolate from the same blood culture

2020 ◽  
Author(s):  
Kyriaki Xanthopoulou ◽  
Julia Wille ◽  
Janine Zweigner ◽  
Kai Lucaßen ◽  
Thorsten Wille ◽  
...  
Keyword(s):  
Blood Culture ◽  
Enterococcus Faecium ◽  
Core Genome ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Vana Gene ◽  
Plasmid Analysis ◽  
Vancomycin Resistant ◽  
Faecium Isolate

Abstract Objectives To characterize two Enterococcus faecium isolates with different resistance phenotypes obtained from the same blood culture. Methods The isolates were identified by MALDI-TOF MS and antimicrobial susceptibility testing (AST) was performed using a VITEK® 2 AST P592 card and Etest. WGS was performed on the MiSeq and MinION sequencer platforms. Core-genome MLST (cgMLST) and seven-loci MLST were performed. Plasmid analysis was performed using S1-PFGE followed by Southern-blot hybridization. Results Both E. faecium isolates were ST203. AST revealed that one was a vancomycin-resistant E. faecium (VREfm) isolate and the other was a vancomycin-susceptible E. faecium (VSEfm) isolate. The VREfm isolate harboured the vanA gene cluster as part of a Tn1546-type transposon encoded on a 49 kb multireplicon (rep1, rep2 and rep7a) plasmid (pAML0157.1). On the same plasmid, ant(6)-Ia, cat-like and erm(B) were encoded. The VSEfm isolate harboured a rep2 plasmid (pAML0158.1), 12 kb in size, which was present in full length as part of pAML0157.1 from the VREfm isolate. The vanA-encoding pAML0157.1 was a chimera of the rep2 pAML0158.1 and a second DNA segment harbouring vanA, ant(6)-Ia, erm(B) and cat-like, as well as the replicons rep1 and rep7a. By cgMLST analysis, the VREfm and VSEfm isolates were identical. Conclusions Our results demonstrate that the VREfm and VSEfm blood culture isolates represented ST203 and were identical. The investigated heterogeneous resistance phenotypes resulted from the acquisition or loss of plasmid segments in the enterococcal isolates. These data illustrate that mobile genetic elements may contribute to the spread of vancomycin resistance among enterococci and to the genotypic and phenotypic variation within clonal isolates.

scholarly journals Characterization of the bla KPC-2 and bla KPC-3 genes and the novel bla KPC-15 gene in Klebsiella pneumoniae

2014 ◽  
Vol 63 (7) ◽  
pp. 981-987 ◽  
Author(s):  
Dongguo Wang ◽  
Wei Hou ◽  
Jiayu Chen ◽  
Yonghua Mou ◽  
Linjun Yang ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Gene Mapping ◽  
Southern Blot ◽  
Blot Hybridization ◽  
Carbapenem Resistance ◽  
Southern Blot Hybridization ◽  
The Novel ◽  
Plasmid Analysis ◽  
High Level

Three Klebsiella pneumoniae isolates exhibiting high-level resistance to carbapenem were analysed by PCR, PFGE, gene mapping, plasmid conjugation and Southern blot hybridization using a bla KPC probe. In addition to the frequently reported bla KPC-2 and bla KPC-3 genes, a novel bla KPC-15 gene was identified in one of the isolates. The results of plasmid analysis and Southern blot hybridization revealed that the three bla KPC genes were located on transferable plasmids exhibiting three different patterns. The patterns A, B and C were observed in the genetic makeup of each individual plasmid, and all three structures contained ISKpn6-like and ISKpn8 transposons. The results of the gene mapping and hybridization experiments performed with the bla KPC probe demonstrated that the plasmids harboured the three genes at approximately the 85.0, 54.0 and 73.0 kb positions. The study concluded that carbapenem resistance in the three isolates was primarily due to the production of carbapenem-hydrolysing β-lactamase.

scholarly journals Characterization of two BHV-4 strains isolated in the Czech Republic

2010 ◽  
Vol 55 (No. 3) ◽  
pp. 106-112 ◽  
Author(s):  
V. Fichtelova ◽  
K. Kovarcik
Keyword(s):  
Reference Strain ◽  
Fragment Size ◽  
Blot Hybridization ◽  
Size Variation ◽  
Southern Blot Hybridization ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
The Czech Republic ◽  
Herpesvirus 4

This study describes the isolation of bovine herpesvirus 4 (BHV-4) from the respiratory tract of animals suffering from respiratory disease. DNA of new isolates, CH and Ni, was cleaved with <I>Bam</I>HI, <I>Eco</I>RI and <I>Hind</I>III in restriction enzyme analysis and the fragments were identified by co-migration with the restriction profile of the reference strain Movar 33/63 cleaved with the appropriate endonuclease. Typical profiles with polyrepetitive DNA (prDNA) fragments were detected. In order to localize the size variation within the obtained digestion fragments, Southern blot hybridization was performed. Differences between the isolates CH, Ni were localized in both the prDNAs and the unique central part of the genome and were restricted to fragment size variation.

scholarly journals Draft Genome Sequence of a Vancomycin-Resistant and Vancomycin-Dependent Enterococcus faecium Isolate

2016 ◽  
Vol 4 (2) ◽  
Author(s):  
Marion Blaschitz ◽  
Sarah Lepuschitz ◽  
Laura Wagner ◽  
Franz Allerberger ◽  
Alexander Indra ◽  
...  
Keyword(s):  
Genome Sequence ◽  
Enterococcus Faecium ◽  
Prolonged Exposure ◽  
Draft Genome ◽  
Vancomycin Resistance ◽  
Draft Genome Sequence ◽  
Nosocomial Pathogens ◽  
Vancomycin Resistant Enterococci ◽  
Vancomycin Resistant ◽  
Faecium Isolate

Vancomycin-resistant enterococci have emerged as major nosocomial pathogens worldwide. While antimicrobial pressure promotes nosocomial colonization with these enterococci, prolonged exposure to vancomycin may foster the transition from vancomycin resistance to vancomycin dependence. Here, we report the draft genome sequence of a vancomycin-dependent Enterococcus faecium isolate showing partial teicoplanin dependence.

scholarly journals Characterization of Vancomycin-Resistant Enterococcus faecium Isolates from the United States and Their Susceptibility In Vitro to Dalfopristin-Quinupristin

1998 ◽  
Vol 42 (5) ◽  
pp. 1088-1092 ◽  
Author(s):  
G. M. Eliopoulos ◽  
C. B. Wennersten ◽  
H. S. Gold ◽  
T. Schülin ◽  
M. Souli ◽  
...  
Keyword(s):  
United States ◽  
Enterococcus Faecium ◽  
Multiple Drug Resistance ◽  
The United States ◽  
Multiple Drug ◽  
Data Set ◽  
Vancomycin Resistant ◽  
High Level

ABSTRACT In the course of clinical studies with the investigational streptogramin antimicrobial dalfopristin-quinupristin, isolates of vancomycin-resistant Enterococcus faecium were referred to our laboratory from across the United States. Seventy-two percent of the strains were of the VanA type, phenotypically and genotypically, while 28% were of the VanB type. High-level resistance to streptomycin or gentamicin was observed in 86 and 81%, respectively, of the VanA strains but in only 69 and 66%, respectively, of the VanB strains. These enterococci were resistant to ampicillin (MIC for 50% of the isolates tested [MIC50] and MIC90, 128 and 256 μg/ml, respectively) and to the other approved agents tested, with the exception of chloramphenicol (MIC90, 8 μg/ml) and novobiocin (MIC90, 1 μg/ml). Considering all of the isolates submitted, dalfopristin-quinupristin inhibited 86.4% of them at concentrations of ≤1 μg/ml and 95.1% of them at ≤2 μg/ml. However, for the data set comprised of only the first isolate submitted for each patient, 94.3% of the strains were inhibited at concentrations of ≤1 μg/ml and 98.9% were inhibited at concentrations of ≤2 μg/ml. Multiple drug resistance was very common among these isolates of vancomycin-resistant E. faecium, while dalfopristin-quinupristin inhibited the majority at concentrations that are likely to be clinically relevant.

scholarly journals Molecular Characterization of Vancomycin-Resistant Enterococcus faecium Isolates from Korea

2005 ◽  
Vol 43 (5) ◽  
pp. 2303-2306 ◽  
Author(s):  
K. S. Ko ◽  
J. Y. Baek ◽  
J.-Y. Lee ◽  
W. S. Oh ◽  
K. R. Peck ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Enterococcus Faecium ◽  
Vancomycin Resistant Enterococcus ◽  
Vancomycin Resistant

scholarly journals Molecular Characterization of Glycopeptide-Resistant Enterococcus faecium Isolates from Portuguese Hospitals

2005 ◽  
Vol 49 (7) ◽  
pp. 3073-3079 ◽  
Author(s):  
Carla Novais ◽  
João C. Sousa ◽  
Teresa M. Coque ◽  
Luísa V. Peixe
Keyword(s):  
Molecular Characterization ◽  
Enterococcus Faecium ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Vancomycin Resistant ◽  
Geographical Locations

ABSTRACT Fifty-one pulsed-field gel electrophoresis types and 17 Tn1546 variants were identified among 101 Enterococcus faecium isolates recovered in three distant Portuguese hospitals. Intra- and interhospital dissemination of specific strains and Tn1546 types was detected, which might largely contribute to the endemicity of vancomycin-resistant E. faecium in Portuguese hospitals, as happened previously in other geographical locations.

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