Aim: To evaluate and compare the efficacy of single-dose versus multiple-dose antibiotic prophylaxis in preventing surgical site infections (SSIs) in orthopedic and ENT surgeries. Materials and Methods: This randomized comparative study enrolled 100 patients undergoing elective orthopedic and ENT surgeries. Participants were divided into two groups: Group 1 (Single Dose) received a single intravenous antibiotic dose 30 minutes before surgery, while Group 2 (Multiple Doses) received the same initial dose followed by additional doses every 12 hours for 48 hours postoperatively. Patients were monitored for SSIs based on CDC criteria, length of hospital stay, the need for additional antibiotics, and adverse effects. Data were analyzed using SPSS version 25.0, with a p-value of <0.05 considered significant. Results: Both groups were demographically comparable. Group 2 had significantly lower SSI rates (4.00% vs. 10.00%, p = 0.043) and shorter hospital stays (3.9 ± 1.1 days vs. 4.6 ± 1.2 days, p = 0.018). Additional antibiotic use was markedly lower in Group 2 (2.00% vs. 10.00%, p = 0.014). Adverse effects were minimal and comparable between groups (p > 0.05). Multiple-dose regimens were particularly effective in reducing SSIs in orthopedic surgeries, with a rate of 3.00% compared to 10.00% in the single-dose group (p = 0.041). Conclusion: Multiple-dose antibiotic prophylaxis is more effective than single-dose regimens in preventing SSIs, especially in high-risk surgeries such as orthopedics. It reduces SSI incidence, shortens hospital stays, and minimizes the need for additional antibiotics, without increasing adverse effects. Tailored prophylaxis strategies based on surgical risk and patient factors are essential for optimizing outcomes.