Coagulase-negative staphylococci (CNS) are among the most frequently isolated bacterial species in clinical microbiology, and most CNS-related infections are hospital acquired. The aim of the present study was to evaluate the antibiotic resistance profiles of CNS isolated in the First Bethune Hospital. Disk diffusion method was used to study the antimicrobial resistance. The data were analyzed by WHONET 5 software according to Clinical and Laboratory Standards Institute (CLSI). The majority of 2484 strains of CNS were collected from blood (925, 37.2%), secretions (652, 26.2%), urine (323, 13.0%) and pus (250, 10.1%). The percentage of methicillin-resistant coagulase-negative Staphylococci (MRCNS) was 80.5%. All the CNS isolates were sensitive to vancomycin and teicoplanin. MRCNS strains were frequently resistant to multiple antibiotics. Antimicrobial resistance of MRCNS was more serious than that of methicillin-sensitive coagulase-negative Staphylococci (MSCNS). These results suggest that surveillance of antimicrobial resistance among CNS is necessary.