OBJECTIVE: To determine the effectiveness of a care bundle, with a novel line maintenance procedure, in reducing the rate of central line-associated bloodstream infection (CLABSI) in the intensive care unit (ICU). DESIGN, PARTICIPANTS AND SETTING: Before-and-after study using CLABSI data reported to the Victorian Healthcare Associated Infection Surveillance System (VICNISS), in adult patients admitted to a tertiary adult ICU in regional Victoria between 1 July 2006 and 30 June 2014. VICNISS-reported CLABSI cases were reviewed for verification. An intervention was implemented in 2009. INTERVENTION: The care bundle introduced in 2009 included a previously established line insertion procedure and a novel line maintenance procedure comprising Biopatch, daily 2% chlorhexidine body wash, daily ICU central line review, and liaison nurse follow-up of central lines. MAIN OUTCOME MEASURES: CLABSI rate (cases per 1000 central line days). RESULTS: The average CLABSI rate fell from 2.2/1000 central line days (peak of 5.2/1000 central line days in quarter 4, 2008) during the pre-intervention period to 0.5/1000 central line days (0/1000 central line days from July 2012 to July 2014) during the post-intervention period. CONCLUSION: Our study suggests that this care bundle, using a novel maintenance procedure, can effectively reduce the CLABSI rate and maintain it at zero out to 2 years.