Objective To investigate effect of Da Vinci robotic surgery on healthcare-associated infection (HAI) and related factors in surgery patients, and provide evidence for prevention and control for HAI in Da Vinci robotic surgery. Methods Da Vinci robotic surgery cases from May 2016 to October 2018 in a hospital were selected, 839 patients who underwent 7 kinds of surgery were selected as case group, 891 patients who were operated in traditio-nal way and met the inclusion criteria during the same period were as control group, retrospective case-control study was performed. Results Urinary surgery, general surgery, and cardiothoracic surgery accounted for 74.26%, 20.50%, and 5.24% of Da Vinci robotic surgery respectively. HAI case rate, surgical site infection rate, length of hospital stay, duration of antimicrobial use, and duration of indwelling catheterization in the radical resection group of rectal cancer were all lower than control group; length of hospital stay and indwelling catheterization in atrial septal defect repair surgery group were both lower than control group; duration of indwelling catheterization in partial nephrectomy case group was lower than control group, difference were all statistically significant (all P<0.05). Total cost of hospitalization and operation in case group were both higher than control group, difference were both statistically significant (both P<0.01). Conclusion Compared with traditional surgical methods, Da Vinci robotic surgery does not increase the incidence of HAI, some surgeries can reduce the occurrence of HAI.