A first national prevalence survey of healthcare-associated infections (HAIs) was conducted in 830 healthcare facilities in 1996; the prevalence rate (PR) of HAIs was 7.6%. The objective of this survey was to estimate the prevalence of HAIs five years after that initial survey. A "point-in-time" prevalence survey was proposed in June 2001 to French public and private hospitals by the Network for the Alert, Investigation, and Surveillance of Nosocomial Infections (RAISIN). 1,533 facilities participated in the survey, and 305,656 patients were included. The participating facilities accounted for 77% of French hospital beds. The prevalence of HIs was 23,024/305,656 (7.5%). 25% of facilities had an HI prevalence rate of 3.2% or higher, and 5% had an HI prevalence rate of 17% or higher. Four sites accounted for 71% of HIs: urinary tract (40%), skin and mucous membranes (11%), respiratory tract (10%), and surgical site (10%). The prevalence of HIs was lower in private facilities than in public facilities (5.3% vs. 7.5%, p < 10⁻⁶). 2,344/3,652 (64%) S. aureus strains were methicillin-resistant; 335/1,984 (17%) P. aeruginosa strains were ceftazidime-resistant. The prevalence of NI was 13% lower (p < 10⁻⁶) in university hospitals and 24% lower (p < 10⁻⁶) in general hospitals in 2001 than in 1996. This study highlights the need to control bacterial resistance in French healthcare facilities. (R.A.)