Human cytomegalovirus (CMV) is a major pathogen in immunocompromised patients. Transmission in this population is known to occur by fomites, but the potential for airborne spread is unknown. In this study, air from the rooms of two immunosuppressed patients with CMV pneumonia and one patient with latent infection was filtered and examined by a polymerase chain reaction assay. CMV-DNA was easily detected in the rooms of the patients with pneumonia and a weak positive signal was detected in the room of the patient with latent CMV infection. This technique permits the detection of aerosolized CMV-DNA and could possibly be adapted to detect other airborne pathogens.