Objective To explore the application effect of counter-example video combined with scenario simulation teaching in the training of aseptic operation for nursing interns in the operating room. Methods The first step of the study was to collect，sort out the data，and summarize the easy mistakes of aseptic technique to make a counter example video. After that，74 nursing interns practiced in the operating room from December 2021 to July 2022 were selected as participants and divided into a control group and an experimental group（37 in each group）. The control group received theoretical lectures，watched positive operation videos，and then demonstrated the operation. On the basis of intervention in the control group，situational simulations and counter-example videos were implemented in the experimental group. After the training，the theoretical assessment results of aseptic technique，the occurrence rate of error prone points and the teaching effect of interns were compared. Besides，the clinical reality assessment was conducted again 21 days later. Results After teaching，the experimental group and the control group scored（90.11±3.94） points，and（86.29±4.45） points in the examination of aseptic technique respectively. In the second assessment of 21 days later，the experimental group and the control group achieved（94.11±1.83） points and（91.90±1.79） points in clinical reality examination respectively. The two examination results of the examination group were higher than that of the control group，with a statistically significant difference（t=3.847，5.133，P<0.05）. The incidence of error prone points in the experimental group was lower than that in the control group in laying sterile instrument tables，using sterile holding forceps，disposable sterile items，taking sterile solutions，and wearing fully covered sterile surgical gowns（P<0.05）. The experimental group scored higher in evaluating the effectiveness of internship teaching than the control group，with a statistically significant difference（P<0.001）. Conclusion Counter-example video combined with situational simulation teaching can effectively improve the technical quality of sterile operation of nursing interns practiced operating room，and helpful in improving the compliance of standardized operation.