Introduction: Since university education and intensive and limited pre-service training do not provide an acceptable level of adequate performance of tasks by operating room nurses, and considering the limitations of traditional training methods in the field of the operating room; This study was conducted to determine the impact of using the electronic education approach based on the web application, levelled, personalized and based on the needs of nurses on the level of their knowledge in the field of educational courses held. Materials and methods: This study is a quasi-experimental single-group multi-center pre-test-post-test during four phases of educational needs assessment, educational content design, web application design for training and evaluation of operating room nurses and determining the effectiveness of web-based training of operating room nurses. The application was carried out based on their knowledge and satisfaction. During it, 36 nurses from the operating rooms of four selected hospitals in Isfahan, Iran, who met the study's criteria, were included by stratified random sampling based on the determined sample size. The collected data were analyzed using descriptive statistical tests (frequency and frequency percentage, mean and standard deviation), analytical tests (paired t, independent t, ANOVA, Pearson) and SPSS software version 16. took Results: The results showed that the average knowledge scores of the nurses in the operating room in general and in the course of principles of moving, transferring and positioning the patient in the operating room, cardiac anatomy and principles of ergonomics in the operating room in the stage before and after the intervention were equivalent. 5.91 ± 3.96 and 13.67 ± 3.77, 6.07 ± 3.42 and 13.38 ± 1.32, 8.72 ± 3.97 and 18.18 ± 1.07, 2.60 ± 1.57 and 9.10 ± 0.73 after the intervention were significantly higher than before the intervention (P<0.001)). Also, the average satisfaction score of nurses was equal to 21.38 ± 5.83, and 22 (64.7) nurses were delighted with the e-learning course. Also, the average score of nurses' satisfaction in each of the training courses on the principles of moving, transferring and positioning the patient in the operating room, cardiac anatomy and principles of ergonomics in the operating room are respectively equal to 18.76±7.15, 23.36±3.82 and 4.88±22.60 respectively. Seven people (53.8 per cent) from the nurses of the training course groups on the principles of patient movement, transfer and positioning in the operating room were delighted they had. Eight people (72.2 per cent) from the cardiac anatomy group and seven (70 per cent) from the nurses of the ergonomic principles group in the operating room were delighted they had. Conclusion: The results of this study showed that the use of an electronic education approach based on the web application, levelled, personalized and based on the needs of nurses, led to the improvement of the knowledge of operating room nurses. Also, operating room nurses were delighted with electronic training courses. E-learning can be used as a complementary educational tool and method for continuous training of operating room nurses in other specialized fields of operating room and surgery.