Background: Central-line–associated bloodstream infections (CLABSIs) are a significant contributor to morbidity and mortality for neonates; they also increased healthcare costs and duration of hospitalization. This population is susceptible to infections because of their undeveloped immune systems, and they require intravenous access until they can tolerate enteral feedings, which for extremely premature infants can take several weeks (if not months) to achieve. Our hospital is a regional-referral teaching hospital with 772 licensed beds. The neonatal intensive care unit (NICU) is a level 3, 35-bed unit where the most critically ill neonates receive care. After a sustained 3-year period of zero CLABSIs, we identified 10 infections between September 2016 through April 2018. Methods: A multidisciplinary team known as the neonatal infection prevention team (NIPT) was reinstated. This team included members from nursing and infection prevention (IP) and from NICU Shared Governance, as well as a neonatal nurse practitioner (NNP) and a neonatologist to review these CLABSIs. Evidence-based practices, policies, and procedures were implemented to help reduce CLABSIs. Nurse educators provided education and training. The infection prevention team reinstated and modified the central-line maintenance and insertion tools to document compliance and to identify any gaps in care. Nurses were expected to document line maintenance once per shift (a.m. and p.m.). All CLABSIs were entered into the CDC NHSN and the hospital’s safety event reporting system, which required follow-up by a clinical manager. The infection prevention team monitored NHSN standardized infection ratios (SIRs) monthly. The SIR is the number of observed events divided by the number predicted (calculated based on national aggregate data). Results: The highest reported quarterly SIR was 1.423, which occurred in the third quarter of 2018 (Fig. 1). Overall compliance with line maintenance protocols was 86% on the morning shift and 89% on the afternoon shift. With implementation of an evidence-based bundle, the NICU had a rolling 12-month SIR of 0.00 as of October 2019. Conclusions: Multidisciplinary team development, implementation of evidence-based bundle elements, and education on catheter care contributed to the long-term success in decreasing CLABSI rates in our NICU. Although this implementation achieved a zero CLABSI rate, we experienced some barriers, including compliance issues with staff not completing the audit tools, staff turnover, and high patient census.