Evidence-based practice (EBP) has gained significant importance in clinical practice worldwide, including in nursing. This study aimed to explore the potential impact of applying a web-based training program on nurses' knowledge, skills, and attitudes regarding EBP. A quasi-experimental pretest-posttest research design was utilized with a purposive sample of 64 professional nurses who agreed to participate. The study took place in different hospitals and primary healthcare centers in the Bisha Governorate, Aseer region, Saudi Arabia. A four-week standardized web-based training program was implemented using an online learning approach. Nurses were provided with an online self-rated data collection tool through the Google Forms platform. The findings indicated a highly significant difference in the total knowledge and EBP skills mean scores of the post-intervention (53.08±15.9) and (66.03±8.95), respectively compared to pre-intervention (P<0.05). Additionally, there was marked improvement in the mean scores of the positive attitude of the training sessions post-intervention compared to pre-intervention. The program was also well-received by the nurses in terms of quality and usability. The program has the potential to enhance nurses' knowledge, skills, and attitudes toward EBP. Therefore, healthcare organizations may consider adopting web-based training as a means of continuing professional education to promote EBP competencies among nurses.