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Background and Importance Central venous catheter (CVC)–related infections are a common complication in paediatric patients, increasing morbidity and mortality and often necessitating device removal. High concentration antimicrobial lock solutions can eradicate intraluminal biofilm.Aim and Objectives This study aimed to evaluate, by the Antimicrobial Stewardship Program (ASP) team, the quality of antimicrobial lock therapy prescriptions and their adherence to the institutional protocol.Material and Methods Retrospective observational study of paediatric inpatients (January 2022–December 2024) including all CVC antimicrobial lock therapy prescriptions (patient ages 0–18). Variables: age, device, lock agent, isolated microorganisms, institutional protocol adherence. Per protocol, vancomycin is first-line for suspected Gram-positives; ciprofloxacin is first-line for Gram-negatives (better biofilm penetration); amikacin is reserved for resistant strains. Also recorded in vitro susceptibility to the lock solution, antimicrobial changes per susceptibility, treatment duration, and discontinuation reasons. Descriptive analysis: absolute counts and percentages.Results A total of 117 prescriptions from 89 patients were included. Port-a-Cath (PAC) predominated (86.32%), followed by Hickman catheters (6.84%) and peripherally inserted central catheters (PICCs) (4.27%). Lock solutions were vancomycin (74.4%), amikacin (18.8%), ciprofloxacin (3.4%), and liposomal amphotericin B (3.4%). The most frequently isolated microorganisms were Staphylococcus epidermidis (41.88%), Klebsiella pneumoniae (8.55%) and Escherichia coli (5.98%). Analysis of protocol adherence showed that 44.4% of cases were infections caused by Gram-positive microorganisms, all treated with vancomycin (reflecting 100% adherence). Gram-negative infections accounted for 14.5% of cases: 88.2% were treated with amikacin and only 5.9% with ciprofloxacin, reflecting low adherence in the management of Gram-negative bacteria. In 90.6% the empirically lock solution was active according to susceptibility testing. Lock replacement after microorganism identification was required in only 5.98% of cases. Mean treatment duration was 7.43 days. Discontinuation reasons: negative microbiological results (48.72%), catheter removal due to clinical deterioration (39.32%) and death (1.71%).Conclusion and Relevance Monitoring key indicators by Antimicrobial Stewardship Program (ASP) teams enables prioritisation of improvement areas, implementation of new therapeutic strategies and surveillance of microbiological resistance profiles to optimise treatment selection in paediatric patients with CVCs. The low use of ciprofloxacin for Gram-negative infections—despite its first-line status in the protocol—highlights a deviation in clinical practice that represents an opportunity for improvement, warranting prioritised educational initiatives within the involved clinical services.Conflict of Interest No conflict of interest