BACKGROUND: Twenty percent of lung donors in our transplant program carry respiratory carbapenem-resistant (CR) gram-negative bacteria (GNB), most commonly CR Acinetobacter baumannii. Universal multiplex panel testing of lung transplant donors was introduced in June 2022 as a strategy to expedite CR-GNB detection and optimize perioperative antibiotic prophylaxis. We herein describe our experiences with this approach. METHODS: Retrospective single-center cohort study including 53 adult patients who underwent lung-only transplantation between June 2022 and December 2023. RESULTS: The most common bacteria identified by the multiplex panel were Staphylococcus aureus (n = 20), A. baumannii (n = 13), Klebsiella pneumoniae (n = 13), and Pseudomonas aeruginosa (n = 10). The panel detected 6/9 A. baumannii, 2/2 CR K. pneumoniae, 1/1 CR P. aeruginosa, and 7/8 methicillin-resistant S. aureus that were grown on conventional cultures, corresponding to negative predictive values of 94%, 100%, 100%, and 98%. Based on panel or culture results, IV tigecycline was administered as prophylaxis in 19% of patients, colistin in 17%, and novel beta-lactams in 15%. CONCLUSION: The multiplex panel rapidly detected donor CR-GNB with a high negative predictive value and resulted in clinical effects of reducing broad-spectrum antibiotic prescriptions and maintaining adequate posttransplant outcomes. Prospective studies with predefined outcomes are needed to compare panel-directed therapy against current standards of care.