Shortcut detection of the vanB gene cluster in enterococci by a duplex real-time PCR assay.
OBJECTIVE
To develop a robust, simple and rapid method for detection of vanB in enterococci.


METHODS
A real-time duplex PCR assay for the simultaneous detection of Enterococcus faecium and vanB resistance genotype in enterococci was developed in conjunction with a simple method for DNA extraction. The assay was tested on 130 fresh plate cultures of clinical isolates of enterococci and other Gram-positive bacteria.


RESULTS
Forty-eight isolates of vanB E. faecium from 32 different patients and three isolates of vanB E. faecalis were detected within 1 hour. All isolates of E. faecium were identified correctly.


CONCLUSIONS
This simple method for the detection of resistance mediated by vanB is a potentially useful method that is suitable for use in the diagnostic microbiology laboratory.
Adams
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Department of Microbiology, Royal Prince Alfred Hospital, Camperdown, New South Wales, Australia. nick.adams@member.rcpa.edu.au
D000900:Anti-Bacterial Agents / Q000494:pharmacology / Q000627:therapeutic use; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D024881:Drug Resistance, Bacterial / Q000187:drug effects; D016984:Enterococcus faecium / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D005798:Genes, Bacterial / Q000235:genetics*; D005838:Genotype; D016908:Gram-Positive Bacterial Infections / Q000175:diagnosis / Q000188:drug therapy / Q000235:genetics; D006801:Humans; D005810:Multigene Family; D016133:Polymerase Chain Reaction / Q000379:methods*; D020713:Vancomycin Resistance / Q000235:genetics
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004269:DNA, Bacterial; C079913:VanB protein, Enterococcus