Shortcut detection of the vanB gene cluster in enterococci by a duplex real-time PCR assay

SummaryAimsTo develop a robust, simple and rapid method for detection of vanB in enterococci.MethodsA 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.ResultsForty-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.ConclusionThis 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.