Objective To develop a training program for nurses to assist in robotic-assisted surgery and explore its application effect. Methods The initial draft of the training program for nurses assisting in robotic-assisted surgery was constructed based on literature review and semi-structured interviews. From February to April 2022，a total of 26 experts were consulted through two rounds of expert consultation to finalize the training program. In application，32 operating room nurses who received training in robotic-assisted surgery cooperation in a tertiary hospital from April to July 2022 were assigned to the control group，while another 32 operating room nurses who received similar training in the same hospital from August to November 2022 were assigned to the experimental group. The control group received routine training，while the experimental group received training based on the training program for nurses assisting in robotic-assisted surgery. The training effects of the two groups were compared afterward. Results The effective recovery rate of the questionnaires for the two rounds of expert consultation was 100%. The expert authority coefficient was 0.933，and the Kendall’s coordination coefficient was 0.261 and 0.312（P<0.001），respectively. The final training program for nurses assisting in robotic-assisted surgery consisted of six modules and 85 sub-items. After the training，the experimental group performed better than the control group in terms of theoretical assessment scores，installation time of the robotic-assisted surgery system’s sterile hood，calibration time for endoscopic operations using the robotic-assisted surgery system，operation time for robotic-assisted surgery system positioning，first-attempt success rate for robotic-assisted surgery system positioning，and assessment scores for preoperative lithotomy position placement using the robotic-assisted surgery system （all P<0.05）. Conclusion The development of the training program for nurses assisting in robotic-assisted surgery is scientific and reliable. In application it is helpful to improve nurses’ knowledge and skills in assisting robotic-assisted surgery and enhance the training effect.