Nursing staff spend more time with patients with pain than any other health staff member. For this reason, the nurse must possess the basic knowledge to identify the presence of pain in patients, to measure its intensity and make the steps necessary for treatment. Therefore, a prospective, descriptive, analytical, and cross-sectional study was conducted to investigate the knowledge and attitudes regarding pediatric pain in two different populations. The questionnaire, Pediatric Nurses Knowledge and Attitudes Survey Regarding Pain (PKNAS), was applied to 111 hospital pediatric nurses and 300 university nursing students. The final scores for pediatric nurses and nursing students were 40.1 ± 7.9 and 40.3 ± 7.5, respectively. None of the sociodemographic variables predicted the scores obtained by the participants (P > 0.05). There was a high correlation between the PKNAS scores of pediatric nurses and nursing students (r = 0.86, P < 0.001). It was observed that the degree of knowledge about pain and its treatment was very low in both groups. Due to this deficiency, pain in children remains inadequately managed, which leads to suffering in this population. It is necessary to increase the continued training in this subject in both areas.