Practical approach to the identification of clinically relevant Enterococcus species

Willey, Barbara M.; Jones, Ronald N.; McGeer, Allison; Witte, Wolfgang; French, Gary; Roberts, Richard B.; Jenkins, Stephen G.; Nadler, Harriette L.; Low, Donald E.

doi:10.1016/s0732-8893(99)00032-2

Cited by 48 publications

(33 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…E. faecalis and E. faecium are also the predominant Enterococcus species in the intestinal microflora of humans and animals and are considered opportunistic pathogens (Willey et al 1999), while E. mundtii is associated with plants and soil and is rarely associated with human infection (Pinto et al 1999). In this study, these three environmentally associated species products that may also be suitable.…”

Section: Discussionmentioning

confidence: 96%

Microbiological evaluation of fecal bacterial composition from surface water through aquifer sand material

Ibekwe

Lyon

2008

Journal of Water and Health

View full text Add to dashboard Cite

When bacterial pathogens from livestock contaminate drinking water supplies, they can cause different forms of gastroenteritis. The objective of this study was to enumerate the concentrations of fecal indicator (Escherichia coli and enterococci) in surface water in order to determine removal efficiency by sand filtration. The concentrations of different indicator bacterial species were determined after running tertiary treated water through two tanks containing aquifer material. Enterococcus faecalis primers targeting the ddl gene and primers for Enterococcus faecium were used to identify the two species in the samples. A PCR assay based on the partial sequence of the b-D-glucoronidase gene (uidA) for specific detection and differentiation of E. coli populations was used to confirm the presence of E. coli after a biochemical test. The biochemical test overestimated the percentage of E. faecium in our samples, but the PCR assay with the ddl gene produced 100% specificity with Enterococcus faecalis. The biochemical test was 91.5% specific in identifying E. coli. The composition of indicator bacteria in Santa Ana River was dominated by intestinal microflora of humans and animals; filtration by aquifer sand material may reduce the transport of indicator bacteria from surface water to groundwater.

show abstract

Section: Discussionmentioning

confidence: 96%

Microbiological evaluation of fecal bacterial composition from surface water through aquifer sand material

Ibekwe

Lyon

2008

Journal of Water and Health

View full text Add to dashboard Cite

show abstract

“…Conventional methods of identification of Enterococcus species are time-consuming (7,32). Errors can happen and are not easily detected in automated identification systems, and supplemental testing is sometimes required for identification (14).…”

Section: Discussionmentioning

confidence: 99%

“…Conventional identification methods, which are based on phenotypic and culture characteristics, require 2 to 3 days to provide results (7,32). Currently, species identification of enterococci in many laboratories relies on automation or rapid kits.…”

mentioning

confidence: 99%

Determination of Enterococcus faecalis groESL Full-Length Sequence and Application for Species Identification

et al. 2001

View full text Add to dashboard Cite

Amplification of the partial Cpn60 (or GroEL) gene segment has been used for identification of many bacteria, including Enterococcus species. To obtain more sequence data from groESL genes of Enterococcus faecalis, the full-length sequence of the E. faecalis groESL genes containing groES (285 bp), spacer (57 bp), and groEL (1,626 bp) was determined. A database search of GenBank revealed that the deduced E. faecalis GroES and GroEL proteins show significant homology to the GroES and GroEL proteins of other bacteria. The GroEL (groEL) of E. faecalis had the highest identity with Streptococcus pneumoniae (81.8% amino acid sequence identity and 73.0% nucleotide sequence identity), followed by Lactococcus zeae, while GroES (groES) had 60.2% (64.6%) identity with Lactobacillus zeae and 58.5% (66.2%) identity with Lactococcus lactis, followed by 57.0% (65.5%) identity with Bacillus subtilis. Based on the groES sequence, an E. faecalis-specific PCR assay was developed, and this PCR assay was positive for all the E. faecalis strains tested. Dot blot hybridization using either groES or groEL as the probe distinguished E. faecalis clearly from other species, indicating that both genes can be used as suitable targets for E. faecalis identification. Moreover, broad-range PCR-restriction fragment length polymorphism of groESL was designed to differentiate eight commonly encountered Enterococcus species. The Enterococcus species of reference strains could be easily differentiated on the basis of restriction patterns produced by HaeIII and RsaI. The DNA-based assays developed in this study provide an alternative to currently used methods of identification for clinically important enterococcal species.

show abstract

“…Although most human enterococcal infections are caused by E. faecalis and E. faecium, various studies have revealed an increase in infections caused by E. durans, E. hirae, E. gallinarum and E. casseliflavus (Baele et al, 2000;Kirschner et al, 2001;Knijff et al, 2001;Willey et al, 1999). There is, therefore, a need for rapid and accurate identification of enterococci at species level, as a means of effective infection control.…”

Section: Introductionmentioning

confidence: 99%

Application of multilocus sequence analysis (MLSA) for rapid identification of Enterococcus species based on rpoA and pheS genes

et al. 2005

View full text Add to dashboard Cite

The aim of this study was to evaluate the use of RNA polymerase a subunit (rpoA) and phenylalanyl-tRNA synthase (pheS) gene sequences as species identification tools for enterococci. Ninety-six representative strains comprising all currently recognized Enterococcus species were examined. rpoA gene sequences generated a robust classification into species groups similar to the one based on 16S rRNA gene sequence analysis. On the other hand, the pheS gene is a fast-evolving clock even better suited for species delineation than the rpoA gene, but not for recognition of species groups within Enterococcus as determined by both rpoA and 16S rRNA genes. All enterococcal species were clearly differentiated on the basis of their rpoA and pheS sequences. Evaluation of intraspecies variation showed that both rpoA and pheS genes have a high degree of homogeneity among strains of the same species. Strains of the same enterococcal species have at least 99 % rpoA and 97 % pheS gene sequence similarity, whereas, different enterococcal species have at maximum 97 % rpoA and 86 % pheS gene sequence similarity. It was concluded that both genes can be used as reliable tools for identification of clinical and environmental species of Enterococcus and are efficient screening methods for the detection of novel species. The sequence data obtained in this study were compared to the available atpA and 16S rRNA gene sequences. The MLSA approach to Enterococcus taxonomy provides portable, highly reproducible data with lower costs for rapid identification of all enterococcal species.

show abstract

Practical approach to the identification of clinically relevant Enterococcus species

Cited by 48 publications

References 23 publications

Microbiological evaluation of fecal bacterial composition from surface water through aquifer sand material

Microbiological evaluation of fecal bacterial composition from surface water through aquifer sand material

Determination of Enterococcus faecalis groESL Full-Length Sequence and Application for Species Identification

Application of multilocus sequence analysis (MLSA) for rapid identification of Enterococcus species based on rpoA and pheS genes

Contact Info

Product

Resources

About