Objective To evaluate the antibacterial effect of lidocaine/prilocaine on Staphylococcus aureus (S.aureus). Methods One hundred and twenty-six S. aureus isolates were collected from patients with skin and soft tissue infection (SSTI). The drug resistance of S.aureus towards 18 antibiotics, including penicillin, cefoxitin, and gentamicin, were measured by the disk diffusion method. Methicillin-resistant S.aureus (MRSA) was screened based on the result of antimicrobial susceptibility test of cefoxitin and mecA gene. The minimum inhibitory concentration (MIC) of vancomycin was detected by E-test. The agar dilution method was adopted to detect the MIC of lidocaine/prilocaine. Results Twenty-three of 126 S. aureus isolates were MRSA. The resistance rates of penicillin and erythromycin were higher and were 86.5% and 46.8%, respectively. No isolates were resistant to teicoplanin, vancomycin, linezolid, and quinupristin/dalfopristin. The MIC range of S.aureus lidocaine/prilocaine towards was (1/1~2/2) mg/mL and MIC50 and MIC90 were both (2/2) mg/mL. Conclusion Lidocaine/prilocaine shows significant antibacterial effect on S.aureus isolated from patients with skin and soft tissue infections. They can stop pain and itch for patients with skin diseases and effectively prevent and inhibit S. aureus infections.