In this study, we aim to estimate the risk of developing clinical multidrug-resistant organism (MDRO) infection with carbapenem-resistant Enterobacterales (CRE), methicillin-resistant Staphylococcus aureus (MRSA), or vancomycin-resistant enterococci (VRE) in colonized patients compared with non-colonized admitted to high-risk areas with a main focus on CRE colonization/infection. Retrospective cohort study conducted at a tertiary care facility. This study included patients enrolled in active surveillance testing (AST) for CRE, MRSA, or VRE during the year 2021. Development of relevant invasive infection within 365 days of the AST result was collected as the primary outcome. The association between MDRO colonization and infection was calculated using the risk ratio. The prevalence of CRE organisms and carbapenemase genes is presented. A total of 19,134 ASTs were included in the analysis (4,919 CRE AST, 8,303 MRSA AST, and 5,912 VRE AST). Patient demographics were similar between colonized and non-colonized groups. Colonization was associated with an increased risk of infection in the 3 cohorts (CRE, MRSA, and VRE), with risk ratios reported as 4.6, 8.2, and 22, respectively. Most patients (88%) develop CRE infection with the same colonizing carbapenemase gene. Oxa-48/NDM Klebsiella pneumoniae was the most common organism detected in CRE infection. The study demonstrated that colonization with CRE, MRSA, or VRE is a risk factor for developing infections caused by the respective bacteria. The high percentage of match between carbapenemase genes detected in colonization and infection indicates that screening results might be used to inform infection management and treatment.