The incidence of tracheal colonization and its association with ventilator-associated pneumonia caused by methicillin-sensitive Staphylococcus aureus (MSSA) was studied prospectively in 530 consecutively admitted mechanically ventilated patients in a general intensive care unit. Furthermore, the clinical spectrum, outcome, and microbiological results of 27 cases of staphylococcal ventilator-associated pneumonia (SVAP) were examined. Ventilator-associated pneumonia was diagnosed by protected specimen brush and/or bronchoalveolar lavage. On admission, 7% of the patients were colonized with MSSA in the trachea. Acquired tracheal colonization was demonstrated in 10% of the patients and occurred less frequently in patients with a hospital stay of > 48 h before ICU admission compared to patients admitted directly to the ICU (6% vs. 15%, p < 0.001). Moreover, colonization was acquired more frequently among trauma and neurological/neurosurgical patients (22%) as compared to surgical and medical patients (7%) (p < 0.0001). Twenty-one patients (4%) developed SVAP, the incident being higher in patients colonized in the trachea with MSSA than in those not colonized (21% vs. 1%), p < 0.00001). Staphylococcal ventilator-associated pneumonia developed more often in trauma and neurological/neurosurgical patients as compared to surgical and medical patients (8% vs. 3%, p < 0.05). Moreover, patients with a hospital stay of < 48 h before admission to the ICU had a higher incidence of SVAP as compared to those with a longer hospital stay before ICU admission (7% vs. 2%, p < 0.01). Crude infection-related mortality was 26%. Preceding colonization with MSSA in the trachea appears to be an important risk factor for the development of SVAP, and patients with a short duration of hospitalization before intensive care unit admission have the highest incidence of ventilator-associated pneumonia caused by MSSA.