title: Development and evaluation of a Quadruplex Taq Man real-time PCR assay for simultaneous detection of clinical isolates of Enterococcus faecalis, Enterococcus faecium and their vanA and vanB genotypes.
abstract: OBJECTIVE
  We developed and evaluated the utility of a quadruplex Taqman real-time PCR assay that allows simultaneous identification of vancomycin-resistant genotypes and clinically relevant enterococci.
  
  
  METHODS
  The specificity of the assay was tested using reference strains of vancomycin-resistant and susceptible enterococci. In total, 193 clinical isolates were identified and subsequently genotyped using a Quadruplex Taqman real-time PCR assay and melting curve analysis. Representative Quadruplex Taqman real-time PCR amplification curve were obtained for Enterococcus faecium, Enterococcus faecalis, vanA-containing E. faecium, vanB-containing E. faecalis.
  
  
  RESULTS
  Phenotypic and genotypic analysis of the isolates gave same results for 82 enterococcal isolates, while in 5 isolates, they were inconsistent. We had three mixed strains, which were detected by the TaqMan real-time PCR assay and could not be identified correctly using phenotypic methods.
  
  
  CONCLUSIONS
  Vancomycin resistant enterococci (VRE) genotyping and identification of clinically relevant enterococci were rapidly and correctly performed using TaqMan real-time multiplex real-time PCR assay.
authors.affiliation: Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.
authors.forename: Taghi
authors.initials: T
authors.lastname: Naserpour Farivar
authors.affiliation: Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Najafipour
authors.affiliation: Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.
authors.forename: Pouran
authors.initials: P
authors.lastname: Johari
authors.affiliation: Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.
authors.forename: Masoumeh
authors.initials: M
authors.lastname: Aslanimehr
authors.affiliation: Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.
authors.forename: Amir
authors.initials: A
authors.lastname: Peymani
authors.affiliation: Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.
authors.forename: Hoasan
authors.initials: H
authors.lastname: Jahani Hashemi
authors.affiliation: Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.
authors.forename: Baman
authors.initials: B
authors.lastname: Mirzaui
keywords: Enterococci; Multiplex TaqMan real-time PCR; Vancomycin
references.citation: Diagn Microbiol Infect Dis. 2003 Jan;45(1):81-4
references.pmid: 12573556
references.citation: Eur J Clin Microbiol Infect Dis. 2012 Mar;31(3):261-5
references.pmid: 21656346
references.citation: J Glob Infect Dis. 2012 Jan;4(1):38-42
references.pmid: 22529626
references.citation: FEMS Immunol Med Microbiol. 2003 Sep 22;38(2):117-26
references.pmid: 13129646
references.citation: Rev Soc Bras Med Trop. 2011 Oct;44(5):631-2
references.pmid: 22031081
references.citation: J Hematol Oncol. 2012 Aug 02;5:46
references.pmid: 22856463
references.citation: Appl Environ Microbiol. 2013 Jan;79(1):196-204
references.pmid: 23087032
references.citation: Fish Shellfish Immunol. 2012 Jul;33(1):99-110
references.pmid: 22554851
references.citation: PLoS One. 2013;8(3):e60589
references.pmid: 23555993
references.citation: Mol Med Rep. 2012 Sep;6(3):488-92
references.pmid: 22710424
references.citation: Indian J Med Microbiol. 2013 Jan-Mar;31(1):47-52
references.pmid: 23508429
references.citation: Appl Microbiol Biotechnol. 2009 Nov;85(1):37-43
references.pmid: 19714325
references.citation: J Glob Infect Dis. 2012 Apr;4(2):106-9
references.pmid: 22754245
references.citation: Clin Microbiol Infect. 2010 Jun;16(6):555-62
references.pmid: 20569266
references.citation: Asian Pac J Cancer Prev. 2012;13(8):3701-4
references.pmid: 23098457
references.citation: Clin Chim Acta. 2007 Sep;384(1-2):1-11
references.pmid: 17689512
references.citation: Environ Sci Technol. 2009 May 1;43(9):3343-8
references.pmid: 19534156
references.citation: Euro Surveill. 2008 Nov 20;13(47):null
references.pmid: 19021959
references.citation: Iran J Med Sci. 2012 Sep;37(3):194-9
references.pmid: 23115452
references.citation: Appl Environ Microbiol. 2012 Feb;78(4):1237-42
references.pmid: 22179252
references.citation: Rev Inst Med Trop Sao Paulo. 1999 Nov-Dec;41(6):339-42
references.pmid: 10671286
references.citation: Infect Immun. 2012 Nov;80(11):3795-803
references.pmid: 22890997
references.citation: J Clin Microbiol. 1999 Nov;37(11):3497-503
references.pmid: 10523541
references.citation: J Pak Med Assoc. 2011 Jun;61(6):586-8
references.pmid: 22204216
references.citation: Biotechnol Lett. 2003 Feb;25(3):261-5
references.pmid: 12882582
references.citation: PLoS One. 2013;8(4):e59491
references.pmid: 23560050
references.citation: Clin Microbiol Infect. 2012 Nov;18(11):E438-45
references.pmid: 22834638
references.citation: Microbiol Immunol. 1996;40(10):773-5
references.pmid: 8981352
references.citation: Infect Control Hosp Epidemiol. 2000 Aug;21(8):520-4
references.pmid: 10968718
references.citation: N Am J Med Sci. 2012 May;4(5):231-4
references.pmid: 22655283
references.citation: J Med Microbiol. 2010 Jan;59(Pt 1):55-64
references.pmid: 19779029