Background: The wide application of antibiotics and antiseptics for patient therapy and medical equipment and surfaces disinfection has resulted in the emergence of resistant microorganisms. Staphylococcus aureus and coagulase-negative Staphylococci (CoNS) are found as a part of the normal resident flora in human so that up to two-thirds of the healthy populations are permanently or transiently colonized by S. aureus and CoNS. Chlorhexidine is an antiseptic agent particularly effective against Gram-positive bacteria. It is widely used for hygienic hand wash to prevent transmission of Staphylococci nosocomial infections. The plasmid-borne qacA/B, qacC, and smr genes confer resistance to cationic antiseptic agents in S. aureus and CoNS. Objectives: The objective of the current study was to characterize the antibiotic resistance and susceptibility to quaternary ammonium compounds (QACs) in methicillin-sensitive S. aureus (MSSA), methicillin-resistant S. aureus (MRSA), methicillin-sensitive coagulase-negative staphylococci (MSCoNS), and methicillin-resistant coagulase-negative Staphylococci (MRCoNS). Methods: In this study, the antibiotic susceptibility and resistance to Chlorhexidine in 120 Staphylococcal strains were evaluated by disc diffusion and Minimum inhibitory concentration (MIC) of Chlorhexidine gluconate (CHG) methods, respectively. The MICs of CHG were determined in triplicate by broth micro-dilution, and the presence of mecA, qacA/B, qacC, and smr genes was examined by PCR assay. Results: Of total 60 S. aureus isolates, 51 (85%) were MRSA, and of 60 CoNS, 7 (11.66%) were MRCoNS. The results showed that the MIC
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