Introduction: The incidence of infections caused by multidrug-resistant Gram-positive bacteria is increasing in the pastyears. The pathogens such as glycopeptid-resistant enterococci, methicillin-resistant Staphylococcus aureus (MRSA), andcoagulase-negative staphylococci (CoNS) have emerged widely and they contribute an increase. The treatment of the patientsinfected with the multidrug-resistant pathogens is extremely difficult. The aim of this study was to determine in vitrosusceptibility of vancomycin-resistant enterococci (VRE) and staphylococci against linezolid that may provide new alternativeto treat.Methods: A total of 80 isolates of staphylococci (30 MRSA, 30 MSSA, and 20 Methicillin-resistant coagulase-negative staphylococci[MRCoNS]) obtained from various clinical specimens and 20 VRE isolates recovered from blood and rectal swabspecimens were sent to our microbiology laboratory. Linezolid susceptibility was determined by disc diffusion methods andE-test for all isolates. In addition, for all isolates, sensitivity to other antibacterials was detected by Kirby–Bauer disc diffusionmethod according to guidelines established by the Clinical and Laboratory Standards Institute.Results: The results showed that all strains were fully susceptible to linezolid (minimal inhibitory concentration [MIC] ≤2μG/ml). The linezolid effectiveness is not different between the MRSA and MSSA strains. MICs were changed for MRSA from0.018 to 2 μg/ml, MSSA from 0.25 to 1.5 μg/ml, and MRCoNS from 0.19 to 1 μg/ml. MICs were changed from 0.38 to 2 μg/ml for VRE strains.Discussion and Conclusion: As a result of the present study, it was decided that linezolid appears to be a good alternativein the treatment of infections caused by Gram-positive bacteria, especially those resistant to glycopeptides or with reducedsensitivity.