A retrospective cohort study was conducted among hospitalized children less than 12 years of age who had Acinetobacter spp. isolated from ≥1 cultures between October 2001 and December 2007 at King Abdulaziz Medical City in Riyadh, Saudi Arabia. Children with multidrug-resistant (MDR) Acinetobacter spp. healthcare-associated infections (HAIs) were compared to children with antimicrobial-susceptible Acinetobacter spp. HAIs and to children colonized with Acinetobacter. Children with MDR Acinetobacter spp. HAIs were older (p = 0.01), more likely to be admitted to an intensive care unit (ICU) (p = 0.06), and had a higher mortality rate (p = 0.02) than colonized children. Children with MDR Acinetobacter spp. HAIs were older than children with antimicrobial-susceptible Acinetobacter spp. HAIs (p = 0.0004), but their mortality rates were similar. Among children with MDR Acinetobacter spp. HAIs, burn injuries were the most common underlying illness. HAIs caused by MDR or susceptible Acinetobacter spp. occurred after prolonged hospitalization, suggesting nosocomial acquisition. Patients infected with MDR Acinetobacter spp. frequently received inappropriate empiric therapy (73.9 %). Further studies are needed in order to identify effective strategies to prevent nosocomial transmission and effective ways of improving patient outcomes.