title: Detection of glycopeptide resistance genes in enterococci by multiplex PCR.
abstract: BACKGROUND
  Vancomycin Resistant Enterococci (VRE) are a major cause of nosocomial infections. There are various phenotypic and genotypic methods of detection of glycopeptide resistance in enterococci. This study utilizes multiplex PCR for reliable detection of various glycopeptides resistance genes in VRE.
  
  
  METHODS
  This study was conducted to detect and to assess the prevalence of vancomycin resistance among enterococci isolates. From October 2011 to June 2013, a total of 96 non-repetitive isolates of enterococci from various clinical samples were analyzed. VRE were identified by Kirby Bauer disc diffusion method with Clinical and Laboratory Standards Institute (CLSI) guidelines. Minimum inhibitory concentration (MIC) of all isolates for vancomycin and teicoplanin was determined by E-test. Multiplex PCR was carried out for all enterococci isolates using six sets of primers.
  
  
  RESULTS
  Out of 96 isolates, 14 (14.6%) were found to be resistant to vancomycin by vancomycin E-test method (MIC ≥32 μg/ml). Out of these 14 isolates, 13 were also resistant to teicoplanin (MIC ≥16 μg/ml). VanA gene was detected in all the 14 isolates by Multiplex PCR. One of the PCR amplicons was sent for sequencing and the sequence received was submitted in the GenBank (GenBank accession no. KF181100).
  
  
  CONCLUSIONS
  Prevalence of VRE in this study was 14.6%. Multiplex PCR is a robust, sensitive and specific technique, which can be used for rapid detection of various glycopeptide resistance genes. Rapid identification of patients infected or colonized with VRE is essential for implementation of appropriate control measures to prevent their spread.
authors.affiliation: Resident, Dept of Microbiology, Armed Forces Medical College, Pune 411040, India.
authors.forename: Puneet
authors.initials: P
authors.lastname: Bhatt
authors.affiliation: Professor & Head, Dept of Microbiology, Armed Forces Medical College, Pune 411040, India.
authors.forename: A K
authors.initials: AK
authors.lastname: Sahni
authors.affiliation: Professor and Head, Dept of Microbiology, All India Institute of Medical Sciences, Bhubaneswar, India.
authors.forename: A K
authors.initials: AK
authors.lastname: Praharaj
authors.affiliation: Associate Professor, Dept of Microbiology, Armed Forces Medical College, Pune 411040, India.
authors.forename: Naveen
authors.initials: N
authors.lastname: Grover
authors.affiliation: Associate Professor, Dept of Microbiology, Armed Forces Medical College, Pune 411040, India.
authors.forename: Mahadevan
authors.initials: M
authors.lastname: Kumar
authors.affiliation: Professor (Microbiology), INHS Asvini, Mumbai, India.
authors.forename: C N
authors.initials: CN
authors.lastname: Chaudhari
authors.affiliation: Resident (Microbiology), All India Institute of Medical Sciences, Bhubaneswar, India.
authors.forename: Atul
authors.initials: A
authors.lastname: Khajuria
keywords: Clinical and Laboratory Standards Institute; GenBank; Minimum inhibitory concentration; Multiplex PCR; Vancomycin resistant enterococci