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Central venous access device-related thrombosis (CRT) is a common complication in hospitalized children. The pediatric nurses' knowledge, attitude, and practice are of great significance for CRT prevention in hospitalized children. The purpose of this study was to elaborate on the level and influencing factors of pediatric nurses' knowledge, attitude, and practice on the prevention related to CRT in hospitalized children. This national cross-sectional study was carried out in China from January 16, 2021, to April 23, 2021. A multi-stage sampling method was adopted, and 1060 pediatric nurses from 21 hospitals participated in this study. The current situation and influencing factors of pediatric nurses' knowledge, attitude, and practice were investigated by descriptive statistics, approximate t test or independent-sample t test, nonparametric Kruskal–Wallis H test, one-way analysis of variance, and multiple linear regression analysis. The relationship among pediatric nurses' knowledge, attitude, and practice was explored by the Pearson correlation analysis. Among all pediatric nurses involved in this study, 25% had insufficient knowledge, 18% had negative attitudes, and 24% had poor behaviors. The main influencing factors on the knowledge, attitude, and practice included the highest education level of pediatric nurses (β = 0.10, P =.001), whether received training related to CRT prevention (β = 0.09–0.14, P <.01), whether CRT-related knowledge was enough for dealing with clinical work (β = 0.18–0.21, P <.001), and the importance of hospitals/departments on children CRT prevention (β = 0.16–0.24, P <.001). There was a positive correlation between knowledge, attitude, and practice (r = 0.24–0.77, P <.01). Conclusion: Pediatric nurses' CRT-prevention knowledge and practice are unsatisfactory, while their preventive attitude toward CRT prevention is optimistic. This study assists the formulation of comprehensive intervention strategies for pediatric nurses in preventing CRT in hospitalized children by hospital-related organizations and nursing managers, so as to improve the participation of pediatric nurses in CRT prevention and reduce the occurrence of CRT for hospitalized children. What is Known: • As a common complication in hospitalized children, CRT can induce many potentially serious complications. • A professional nursing team is an important prerequisite for reducing CRT incidence. What is New: • The levels of pediatric nurses' knowledge and practice are not satisfactory, while pediatric nurses' preventive attitude toward CRT prevention is optimistic. • Hospital-related organizations and nursing managers should highlight the importance of CRT prevention and encourage pediatric nurses to improve their expertise and strengthen the training related to CRT prevention. [ABSTRACT FROM AUTHOR]