After isolation of multiresistant (MR) Pseudomonas aeruginosa from 3 hospitalized patients in a paediatric intensive care unit (PICU), a prospective surveillance programme was established to detect infected and/or colonized patients in the hospital. Isolates were examined by means of outer membrane protein (OMP) profiles, serotyping and DNA genomic analysis using pulsed-field gel electrophoresis (PFGE). Fifty-five P. aeruginosa strains were isolated from 23 hospitalized patients during September and October 1997. The median hospital stay before isolation of P. aeruginosa was 8 d. PFGE demonstrated that the same clone infected 14 patients, 4 of whom were not hospitalized in the PICU. Susceptibility patterns and OMP profiles correlated with PFGE results in 37.8% and 36.4% of cases, respectively. Serotype O11 correlated with pattern A in 77% of cases and serotype O4 correlated with unrelated strains in 75% of cases but did not discriminate between outbreak and unrelated isolates. Extensive investigation of cultures failed to identify a reservoir of P. aeruginosa. PFGE was superior to OMP analysis and serotyping for discriminating between strains. The possible mode of acquisition for most of the patients infected with the same clone was cross-contamination.