Concomitant lung colonization by Aspergillus fumigatus and Stenotrophomonas maltophilia was reported mainly in patients with cystic fibrosis (CF) and immunocompromised patients. The aim of the study was to assess the frequency of co-culture of A. fumigatus and S. maltophilia in respiratory samples of hospitalized patients, and to determine its associated factors. Between 2007 and 2011, all patients who had A. fumigatus in their respiratory samples were retrospectively enrolled in the study. Their clinical and laboratory data, including the presence of S. maltophilia in a respiratory sample, were collected within the same month. Of the 257 enrolled patients (372 respiratory samples), 71 % were immunocompromised and 32 % had chronic respiratory disease. S. maltophilia was isolated within the same month in 20 patients (7.8 %). In the univariate analysis, factors associated with concomitant culture of A. fumigatus and S. maltophilia were liver disease (P = 0.009), orotracheal intubation (P = 0.001), ventilator-associated pneumonia (P = 0.006), central venous catheter (P = 0.003), parenteral nutrition (P = 0.008) and culture of Pseudomonas aeruginosa in respiratory samples (P = 0.002). In the multivariate analysis, the simultaneous presence of P. aeruginosa in the respiratory tract (odds ratio (OR) = 3.19, 95 % confidence interval (CI) 1.11–9.14, P = 0.031), liver disease (OR = 3.92, 95 % CI 1.32–11.62, P = 0.014) and orotracheal intubation (OR = 3.42, 95 % CI 1.17–9.96, P = 0.024) were independently associated with the co-culture of S. maltophilia and A. fumigatus. Factors independently associated with the concomitant culture of A. fumigatus and S. maltophilia were identified. These results support a future prospective study focusing on liver disease and its complications.