BACKGROUND The rising prevalence of carbapenem-resistant Acinetobacter baumannii (CRAB) in neonatal intensive care units (NICUs) represents an escalating challenge in healthcare settings, particularly in managing hospital-acquired infections (HAIs). Studies across various World Health Organization regions have documented a significant incidence of CRAB-related HAIs, with rates as high as 41.7 cases per 1000 patients in ICUs, accounting for 13.6% of all HAIs. These infections pose a doubled mortality risk compared to infections with carbapenem-susceptible Acinetobacter baumannii . A particularly concerning aspect of CRAB colonization is its asymptomatic nature, enabling its transmission through healthcare workers (HCWs) or the NICU environment to vulnerable neonates with developing immune systems. AIM To explore the prevalence of CRAB colonization in NICUs, focusing on neonates, healthcare workers, and the environmental samples, to enhance epidemiological understanding and inform targeted interventions. METHODS We conducted according to PRISMA 2020 checklist guidelines, a comprehensive literature search across multiple databases including MEDLINE (Ovid), EMBASE (Ovid), Global Health (Ovid), Web of Science, and Global Index Medicus. Studies were selected based on predetermined criteria, primarily involving neonates, HCWs, and environmental swabs, using culture or molecular methods to detect CRAB colonization. We excluded studies that did not specifically focus on NICUs, were duplicates, or lacked necessary data. The study selection and quality assessment were conducted independently by two reviewers. Data extraction involved collecting comprehensive details about each study. Our statistical analysis used a random-effects model to calculate the pooled prevalence and confidence intervals, stratifying results by regional location. We assessed study heterogeneity using Cochran's Q statistic and I ² statistic, with regression tests employed to evaluate potential publication bias. RESULTS We analyzed 737 records from five databases, ultimately including 13 studies from ten countries. For neonates, the pooled prevalence was 4.8% (95%CI: 1.1% to 10.5%) with the highest rates observed in South-East Asia (10.5%; 95%CI: 2.4% to 23.3%). Among HCWs, a single Indian study reported a 3.3% prevalence. Environmental samples showed a prevalence of 2.3% (95%CI: 0% to 9.3%), with the highest rates in South-East Asia (10%; 95%CI: 4.2% to 17.7%). Significant heterogeneity was found across studies, and no publication bias was detected. CONCLUSION This systematic review highlights a significant prevalence of CRAB colonization in neonates across various regions, particularly in South-East Asia, contrasting with lower rates in high-income countries. The study reveals a gap in research on HCWs colonization, with only a single study from India reporting moderate prevalence. Environmental samples indicate moderate levels of CRAB contamination, again higher in South-East Asia. These findings underscore the need for more extensive and focused research on CRAB colonization in NICUs, including exploring the roles of HCWs and the environment in transmission, understanding antimicrobial resistance patterns, and developing effective prevention measures.