Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 6 5 2014 10 15 335 340 EN Taghi Naserpour-Farivar Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran. Reza Najafipour Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran. Pouran Johari Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran. Masoumeh Aslanimehr Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran. Amir Peymani Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran. Hoasan Jahani Hashemi Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran. Baman Mirzaui Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran. 2015 10 16 Background and Objectives: 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. Materials and 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. Conclusion: 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. https://ijm.tums.ac.ir/index.php/ijm/article/view/375 https://ijm.tums.ac.ir/index.php/ijm/article/download/375/152