Objectives: Surgical site infections (SSIs) remain a common and global problem that contributes to significant morbidity and mortality, prolongs hospital stay, and consequently increases health-care costs. Bacterial resistances pose a challenge and complicated the SSI treatment. This study aimed to evaluate the bacterial profile and antimicrobial susceptibility patterns of isolates among patients diagnosed with surgical site infection. Methods: Patients who underwent either elective or emergency surgical procedures were enrolled in this study. For those who developed surgical site infections, specimens from the surgical site were collected and processed at the microbiology laboratory at the tertiary health-care center, Ahmedabad. Results: Out of the 5003 patients, 34 developed SSI. In our study, 37 bacteria were isolated from 34 samples subjected to culture. Thirty-two samples showed monomicrobial growth and two showed polymicrobial growth. Among them, 14 (38%) were Gram-positive cocci, out of which 12 (85.7%) were Staphylococcus aureus and 2 (14.3%) were Enterococci spp. and 23 (62%) were Gram-negative bacilli, there were 21 (91.3%) enterobacteriaceae, out of which 5 (23.8%) were Escherichia coli, 1 (4.8%) were Proteus mirabilis, 15 (71.4%) Klebsiella spp. and 02 (8.7%) non-fermenters of which Pseudomonas aeruginosa and Acinetobacter baumannii. Conclusions: Gram-negative bacteria were the most dominant isolates from surgical sites in the study area. Among them, Klebsiella spp. were the most common bacteria isolated from surgical site specimens. In our study, there is high antibiotic resistance observed which raises serious concerns and mandates strict antibiotic policy as well as antimicrobial stewardship.