Aim: Surgical site infections (SSIs) are a major cause of morbidity and prolonged hospital stays, often complicated by antimicrobial resistance. This study aims to determine the prevalence of SSIs, identify the causative pathogens, and analyze their antibiotic susceptibility patterns in a tertiary care hospital. Material and Methods: This prospective study was conducted in the Department of Microbiology at K.D. Medical College Hospital and Research Centre (KDMCH), Mathura, Uttar Pradesh. A total of 120 patients with SSIs were included, and samples were collected using sterile swabs or aspirates. The isolates were identified using standard microbiological techniques and tested for antibiotic susceptibility using the Kirby-Bauer disc diffusion method as per Clinical and Laboratory Standards Institute (CLSI) guidelines. Resistance patterns, including methicillin-resistant Staphylococcus aureus (MRSA), extended-spectrum beta-lactamase (ESBL), and carbapenemase production, were also analyzed. Results: The highest incidence of SSIs was observed in the 41-60 years age group (37.50%), with general surgical procedures being the most affected (41.67%). Staphylococcus aureus (33.33%) and Escherichia coli (25.00%) were the predominant isolates. Antibiotic susceptibility testing revealed high resistance to amoxicillin (70.00%) and ceftriaxone (50.00%), while meropenem exhibited the highest sensitivity (90.00%). Multidrug-resistant organisms were identified, including MRSA (12.50%), ESBL producers (16.67%), and carbapenemase-producing bacteria (8.33%). Conclusion: The study highlights the burden of SSIs and the rising challenge of antimicrobial resistance, particularly among Staphylococcus aureus and Escherichia coli. High resistance rates to commonly used antibiotics necessitate the implementation of antimicrobial stewardship programs and stringent infection control measures. Continuous surveillance of pathogen resistance patterns is crucial for guiding effective empirical therapy and improving patient outcomes.