Varicella-zoster virus (VZV) is a highly contagious infectious agent that causes outbreaks in institutional settings. Transmission of VZV is felt to occur following direct contact with an infected individual and by aerosol spread. To document the aerosolization ofVZV, a polymerase chain reaction (PCR) assay was used to detect VZV DNA in air samples obtained from hospital rooms of patients with active VZV infection. VZV DNA was detected in 64 (82%) of 78 air samples from rooms housing patients with active varicella and 9 (70%) of 13 samples from rooms of patients with herpes zoster. VZV was detected 1.2–5.5 m from patients' beds and for 1–6 days following onset of rash. On some occasions, VZV DNA could be detected outside the hospital isolation rooms housing patients. This PCR-based method allows the detection and semiquantitation of VZV aerosolization and can be a useful tool for monitoring efforts to control VZV aerosols in the environment.