Teicoplanin is an antibiotic used to treat severe Gram-positive infections, especially those caused by methicillin-resistant Staphylococcus aureus(MRSA). In this study, we aimed to evaluate the pattern of teicoplanin rational prescribing to identify the factors which affected rational utilization. In addition, the teicoplanin minimum inhibitory concentration (MIC) was assessed in randomly selected isolates. Methods: In this descriptive-analytical prospective study, a total of 256 patients were randomly selected to evaluate the pattern of teicoplanin use. The required data were gathered to assess the appropriateness of teicoplanin usage. Also, 100 teicoplanin Etests were used for measuring the MIC. Findings: The results showed that the appropriateness rate of teicoplanin usage was 21.9%. The mean MIC was 2.24 ± 5.47 mg/L for the MRSA cultures (33 cultures), including 32 sensitive cultures (97%). In addition, the mean MIC was 28.71 ± 8.29 mg/L for the vancomycin-resistant enterococci (VRE) cultures (67 cultures), including five sensitive cultures (7.5%). Moreover, the analysis revealed that only the hospitalization ward was statistically significantly related to irrational usage (P = 0.014). Conclusion: The high prevalence of the inappropriate use of teicoplanin will lead to the development of antimicrobial resistance. Furthermore, the high rate of VRE cultures resistant to teicoplanin proves that teicoplanin has no advantage over vancomycin for treating VRE infections. Finally, we recommend guidelines' development for the appropriate administration of teicoplanin.