Detection of Vancomycin-resistant Enterococci using Multiplex Real-time PCR Assay and Melting Curve Analysis / 대한진단검사의학회지
The Korean Journal of Laboratory Medicine
;
: 138-146, 2010.
Article
in Korean
| WPRIM
| ID: wpr-151628
ABSTRACT
BACKGROUND:
We developed and evaluated the utility of a multiplex real-time PCR assay that uses melting curve analysis and allows simultaneous identification of vancomycin-resistant genotypes and clinically relevant enterococci.METHODS:
The specificity of the assay was tested using 4 reference strains of vancomycin-resistant enterococci (VRE) and 2 reference strains of vancomycin-susceptible enterococci. Ninety-three clinical isolates of enterococci with different glycopeptide-resistant phenotypes were genotyped and identified using a multiplex real-time PCR assay and melting curve analysis.RESULTS:
Representative melting curves were obtained for Enterococcus faecium, Enterococcus faecalis, vanA-containing E. faecium, vanB-containing E. faecalis, Enterococcus gallinarum, and Enterococcus casseliflavus. Phenotypic and genotypic analysis of the isolates revealed same results for 82 enterococcal isolates, while in 4 isolates, the glycopeptide-resistant phenotypes were inconsistent with the glycopeptide-resistant genotypes and in the 4 other isolates, species could not be accurately identified. Three isolates with mixed strains, which were detected by the PCR assay, could not be correctly identified using phenotypic methods.CONCLUSIONS:
VRE genotyping and identification of clinically relevant enterococci were rapidly and correctly performed using multiplex real-time PCR assay and melting curve analysis.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Peptide Synthases
/
Phenotype
/
Bacterial Proteins
/
DNA, Bacterial
/
Polymerase Chain Reaction
/
Enterococcus faecium
/
Enterococcus
/
Enterococcus faecalis
/
Carbon-Oxygen Ligases
/
Vancomycin Resistance
Type of study:
Diagnostic study
Language:
Korean
Journal:
The Korean Journal of Laboratory Medicine
Year:
2010
Type:
Article
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