Species Specific Detection of Methicillin-resistant Staphylococcus aureus (MRSA) by Multiplex PCR

Document Type : Original Article(s)

Authors

1 PharmD Student, Department of Biotechnology, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran

2 Assistant Professor, Department of Biotechnology, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran

3 Department of Microbiology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran

4 MSc Student, Department of Microbiology, Islamic Azad University, Science and Research Branch, Arak, Iran

Abstract

Introduction: Staphylococcus aureus is one of the pathogens that is a major agent of infectious diseases. So detection, control and treatment of this pathogen is very important. The objective of this study is setting up a fast method to detect methicillin resistant Staphylococcus aureus (MRSA) for better management of diseases related to this pathogen.Materials and methods: In this study 225 sample collected form staff of University hospitals of Zanjan province. After detection and confirmation of bacteria by conventional and biochemical methods, antimicrobial susceptibility was tested by disk diffusion (Kirby-Bauer) method. After DNA extraction by boiling and phenol – chloroform method, Sa442, mecA and femA genes were detected by multiplex PCR and PCR products were analyzed by electrophoresis (70V, 30 min) in gels composed of 1.5% (w/v) agarose stained with SyberGreen.Result: 208 out of 225 samples made colony in bacterial culture medium. 203/208 isolates were gram positive; 201/208 isolates were catalase positive and 71/208 isolates were coagulase positive. Most of the isolates were resistant to oxacillin and penicillin. By optimization of PCR conditions, Sa442, mecA and femA genes were detected by multiplex PCR simultaneously in single reaction tube.Discussion: Multiplex PCR can detect rapidly and selectively methicillin resistant Staphylococcus aureus in comparison with disk diffusion method.