Objective To determine the epidemiological, clinical and microbiological characteristics, of patients with post-traumatic osteomyelitis of extremity fractures, and provide evidence-based guidelines for early diagnosis and treatment, including empiric antibiotic therapy. Methods Human subject research was performed using institutional review board approved protocols. A retrospective chart review was conducted on 5,368 patients diagnosed with extremity traumatic fractures from January 1, 2012 to December 31, 2015, to identify osteomyelitis patients. Records from the Microbiology Department were reviewed, and patients with a positive wound culture, or bone biopsy culture, were selected for the study. Microbial suceptability was determined by the M-100-S22 protocol (Clinical & Laboratory Standards Institute? (CLSI) 2012 USA). Additional clinical information, including data on patients' baseline epidemiological, clinical, and microbiological records was collected from all available charts, and reviewed using a designed protocol. Results 84 (1.56%) patients were diagnosed with osteomyelitis based on a positive culture result. The most prevalent comorbidities in these patients were compartment syndrome, diabetes and hypertension. The most commonly involved infected site was the tibia-fibula (47.62%). 66 (78.57%) of these cases were monomicrobial, and 18 cases (21.43%) were polymicrobial. The infections were predominantly caused by Gram-positive bacteria (56, 53.85%). The most common Gram-positive bacteria were Staphylococcus aureus (39 cases, 37.50%) and S. epidermidis (6 cases, 5.77%), which were sensitive to ampicillin, synercid/ dalfopristin, linezolid, tigecycline, macrodantin, and vancomycin. S. aureus was the most common pathogen in both monomicrobial and polymicrobial cases. All 17 cases of MRSA infection were sensitive to Imezolid, ampicillin, synercid/ dalfopristin, linezolid, tigecycline, furadantin, piperacillin/yaz, rifampicin, and vancomycin, respectively. The most common Gram-negative bacteria were E. coli (16 cases, 15.38%) and Enterobacter cloacae (11 cases, 10.58%), which were sensitive to thienamycin. Conclusions In this study, the overall rate of post-traumatic osteomyelitis of limb fractures (1.56%) is lower than the national average rate (2.6-7.8%), for major medical centers in China. The main medical comorbidities were compartment syndrome, diabetes mellitus and hypertension. The most common infection was monomicrobial in lower extremities. S. aureus was the most common pathogen, which presented in 39 (37.50%) cases, and 17 of these (43.59%) were caused by MRSA. These findings can guide empiric antibiotic therapy in Southwest China for osteomyelitis in patients with traumatic limb fractures.