Background: Post-operative wound infections (POWIs) are a leading cause of morbidity and increase in healthcare costs, especially in low resource settings. The rising prevalence of antibiotic resistance among pathogens also increases the complexity of treatment strategies, highlighting the importance of local surveillance of microbiological and resistance patterns. This study aims to find out the antibiotic susceptibility patterns of gram positive and gram negative isolates from the post-operative wound infections from women undergoing obstetric and gynecological surgeries. Methods: A cross-sectional study was conducted at Department of Obstetrics and Gynecology, Dhaka Medical College Hospital, Dhaka from October 2019 to September 2021. A total of 200 women with post-operative wound infections were purposively selected. Among them, 134 had culture-positive findings. Bacteriological analysis and antibiotic susceptibility testing were performed on isolates following standard microbiological methods. Results: Among the 134 culture-positive cases, 33.6% of isolates were gram-positive, with Staphylococcus aureus being predominant. Gram-negative bacteria accounted for 66.4% of isolates, with E. coli and Pseudomonas aeruginosa equally prevalent (20.9%). Gram-positive isolates showed high sensitivity to piperacillin-tazobactam (91.1%) and sulbactam (53.3%) but were resistant to ceftriaxone (48.9%) and cefixime (51.1%). Gram-negative bacteria exhibited sensitivity to amikacin and meropenem but demonstrated widespread resistance to ceftriaxone and doxycycline. Conclusion: This study describes the diverse microbiological profile and high prevalence of antibiotic resistance in POWIs. Novel antibiotic therapy and strict infection control measures are essential to curb the burden of these infections.