Indoor microbial communities play an important role in everyday human health, especially in the intensive care units (ICUs) of hospitals. We used amplicon pyrosequencing to study the ICU microbiome and were able to detect diverse sequences, in comparison to the currently used standard cultivation technique that only detected 2.5% of the total bacterial diversity. The phylogenetic spectrum combined species associated with the outside environment, taxa closely related to potential human pathogens, and beneficials as well as included 7 phyla and 76 genera. In addition, Propionibacterium spp., Pseudomonas spp., and Burkholderia spp. were identified as important sources of infections. Despite significantly different bacterial area profiles for floors, medical devices, and workplaces, similarities by network analyses and strains with identical molecular fingerprints were detected. This information will allow for new assessment of public health risks in ICUs, help create new sanitation protocols, and further our understanding of the development of hospital-acquired infections.