OBJECTIVE: According to the importance of evaluating the antimicrobial resistance pattern in the management of nosocomial infections (NIs), we decided to investigate the prevalence of antimicrobial resistance in Chamran Heart Hospital. METHODS: This retrospective cross-sectional observational study was performed for 6 months from February to July 2022 at Shahid Chamran Hospital of Isfahan, Iran. All hospitalized patients with any NIs were eligible for the study. Clinical specimens were obtained from patients with NIs. All specimens underwent microbial culture, and if bacterial growth developed, differential tests were performed. Antibiotic susceptibility testing also was performed per the standards of Clinical and Laboratory Standards Institute, 2022. FINDINGS: Out of 201 examined samples, urinary infection (34.83%), pneumonia (27.86%), and sepsis (13.43%) were reported to be the most prevalent infections. Among Gram-negatives (76.12%), spp. (26.37%), (24.87%), and spp. (11.44%) were the most common pathogens. About 54.9% of spp., 33.3% of . , and 45.45% of spp. were resistant to carbapenems. About 1.88% and 15% of spp. were identified as pan-drug-resistant bacteria and extensively drug-resistant (XDR), respectively. In addition, 4.34% of spp. were identified as XDR. Among Gram-positives (23.88%), spp. (8.95%) was identified as the most common pathogen, and the prevalence of methicillin-resistant (MRSA) and vancomycin-resistant (VRE) was 11.11% and 61.11%, respectively. CONCLUSION: In our study, carbapenem-resistant Enterobacteriaceae accounts for about 50% of all NIs. Moreover, despite the low prevalence of MRSA, VRE was reported to be high in our center when compared with other studies.