A PCR assay for identification of Enterococcus faecium

Cheng, S; McCleskey, F K; Gress, M J; Petroziello, Joseph M.; Liu, R; Namdari, Hassan; Beninga, Kathleen K.; Salmen, A; DelVecchio, Vito G.

doi:10.1128/jcm.35.5.1248-1250.1997

Cited by 102 publications

(45 citation statements)

References 19 publications

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“…Enterococcus faecium accounts for 5-16% of enterococci isolated from human infections and is second only to Ent. faecalis as the most common Enterococcus species associated with nosocomial infections (Cheng et al 1997). Devriese et al (1987) frequently isolated Ent.…”

Section: Discussionmentioning

confidence: 99%

Characterization of enterococci populations collected from a subsurface flow constructed wetland

Graves

Weaver

2010

Journal of Applied Microbiology

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Aims: The aim of this study was to identify and characterize the population of Enterococcus sp. in domestic wastewater as it flows through a constructed wetland. Methods and Results: Four hundred and eighty‐four Enterococcus isolates were collected from the inlet, various sites within and from the outlet of a plastic lined constructed wetland in College Station, TX. The wetland treated septic tank effluent that passed sequentially through two 1·89 m3 septic tanks and a 1·89 m3 pump tank allowing 48 l doses at a 24 l min−1 rate. The Enterococcus isolates were identified to species using the commercial Biolog system. The 484 Enterococcus isolates were comprised of ten different species, including Enterococcus faecalis (30·6%), Enterococcus pseudoavium (24·0%), Enterococcus casseliflavus (12·8%), Enterococcus faecium (11·2%), Enterococcus mundtii (7·9%), Enterococcus gallinarum (6·2%), Enterococcus dispar (3·7%), Enterococcus hirae (2·1%), Enterococcus durans and Enterococcus flavescens both 0·8%. Of the 88 isolates collected from the inlet, only 9·1% of the isolates were identified as Ent. faecalis and Ent. pseudoavium (36·4%) was identified as the predominant species. Whereas of the 74 isolates collected from the outlet, the predominant species were identified as Ent. faecalis (29·7%). Species identification varied among sites within the wetland, but often Ent. faecalis was the predominant species. Conclusions: Our data suggest that while Ent. faecalis is the predominant species of Enterococcus found in domestic wastewater, the populations may shift during treatment as the wastewater flows through the constructed wetland. Significance and Impact of the Study: We found that shifts in Enterococcus species composition occurred during domestic wastewater treatment. This has implications for the identification of faecal pollution based on the presence of specific bacterial types associated with domestic wastewater.

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Section: Discussionmentioning

confidence: 99%

Characterization of enterococci populations collected from a subsurface flow constructed wetland

Graves

Weaver

2010

Journal of Applied Microbiology

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“…It was not, however, effective against Gram-negative or Leuconostoc strains when assayed on MRS plates. The strain was classified as belonging to E. faecium on the basis of its morphological, biochemical and genetic characteristics together with its sugar fermentation patterns, reliable criteria for distinguishing among Enterococcus species (Mundt 1986;Devriese et al 1991;Cheng et al 1997).…”

Section: Discussionmentioning

confidence: 99%

“…Finally, identification was genotypically confirmed by using speciesspecific PCR amplification. To this end, total DNA from strain F58 obtained using the method of Mora et al (2000) was used as a template together with the specific primers EM1A: cccggctcaaccggg and EM1B: ctctagagtggtcaa described by Cheng et al (1997) (synthesized by Amersham Pharmacia Biotech, Barcelona, Spain). The annealing temperatures assayed were 55, 58 and 62°C.…”

Section: Taxonomic Identificationmentioning

confidence: 99%

“…1 Identification of F58 strain by using species-specific PCR analysis. Amplification was carried out with EM1A and EM1B primers according to Cheng et al (1997) using annealing temperature of 58°C. Lanes 1-4: blank, Enterococcus faecalis A-48-32 (negative control), F58 strain and k-DNA (standard of molecular weight) purity of the samples (Fig.…”

Section: Purification Molecular Mass Determination and Partial Aminomentioning

confidence: 99%

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Enterococcus faeciumF58, a bacteriocinogenic strain naturally occurring in Jben, a soft, farmhouse goat's cheese made in Morocco

Achemchem

Martínez‐Bueno

Abrini

et al. 2005

Journal of Applied Microbiology

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Aims: Characterization of Ent F-58 produced by Enterococcus faecium strain F58 isolated from Jben, a soft, farmhouse goat's cheese manufactured without starter cultures. Methods and Results: E. faecium strain F58 was isolated because of its broad inhibitory spectrum, including activity against food-borne pathogenic and spoilage bacteria. The antimicrobial substance was produced during the growth phase, with maximum production after 16-20 h of incubation at 30°C, and was stable over a wide pH range (4-8) and at high temperatures (5 min at 100°C). The enterocin was purified to homogeneity using cation exchange and hydrophobic interaction on C-18 and reverse-phase high-performance liquid chromatography. The activity was eluted as two individual active fractions (F-58A and F-58B) and matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry analysis showed masses of 5210AE5 and 5234AE3 Da respectively. Both peptides were partially sequenced by Edman degradation, and amino-acid sequencing revealed high similarity with enterocin L50 (I). PCR-amplified fragments containing the structural genes for F-58 A and B were located in a 22-kb plasmid harboured by this strain. We verified that it also holds the structural gene for P-like enterocin. Conclusion: E. faecium strain F58 from Jben cheese, a producer of enterocin L50, exerts an inhibitory effect against strains of genera such as Listeria, Staphylococcus, Clostridium, Brochothrix and Bacillus. Enterocin was characterized according to its functional and biological properties, purification to homogeneity and an analysis of its amino acid and genetic sequences. Significance and Impact of the Study: E. faecium strain F58 is a newly discovered producer of enterocin L50, the biotechnological characteristics of which indicate its potential for application as a protective agent against pathogens and spoilage bacteria in foods.

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“…Species identification in routine and clinical laboratories uses mainly phenotypic methods (Devriese et al 1996). Nevertheless these methods are often unreliable and can take several days (Cheng et al 1997).…”

Section: Introductionmentioning

confidence: 99%

Genetic diversity and safety aspects of enterococci from slightly fermented sausages

et al. 2005

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Aims: To determine the biodiversity of enterococci from slightly fermented sausages (chorizo and fuet) at species and strain level by molecular typing, while considering their safety aspects. Methods and Results: Species-specific PCR and partial sequencing of 16S rRNA and sodA genes were used to identify enterococcal population. Enterococcus faecium was the most frequently isolated species followed by E. faecalis, E. hirae and E. durans. Randomly amplified polymorphic DNA (RAPD)-PCR revealed species-specific clusters and allowed strain typing. Sixty strains of 106 isolates exhibited different RAPD profiles indicating a high genetic variability. All the E. faecalis strains carried virulence genes (efaAfs, esp, agg and gelE) and all E. faecium isolates carried efaAfm gene. Enterococcus faecalis showed higher antibiotic resistance than the other species. Only one E. faecium strain showed vanA genotype (high-level resistance to glycopeptides) and E. gallinarum and E. casseliflavus/flavescens isolates showed vanC1 and vanC2/C3 genotypes (low-level resistance only to vancomycin) respectively. Conclusions: E. faecalis has been mainly associated with virulence factors and antimicrobial multi-resistance and, although potential risk for human health is low, the presence of this species in slightly fermented sausages should be avoided to obtain high quality products. Significance and Impact of the Study: The enterococcal population of slightly fermented sausages has been thoroughly characterized. Several relevant safety aspects have been revealed.

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A PCR assay for identification of Enterococcus faecium

Cited by 102 publications

References 19 publications

Characterization of enterococci populations collected from a subsurface flow constructed wetland

Characterization of enterococci populations collected from a subsurface flow constructed wetland

Enterococcus faeciumF58, a bacteriocinogenic strain naturally occurring in Jben, a soft, farmhouse goat's cheese made in Morocco

Genetic diversity and safety aspects of enterococci from slightly fermented sausages

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