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To test the hypothesis that a considerable number of preterm infants acquire respiratory syncytial virus (RSV) within the hospital during the postnatal stay, a prospective epidemiologic survey was performed. Nasopharyngeal swabs were taken twice weekly for a period of 8 weeks from preterm infants, medical/nursing staff, and parents during the peak of RSV season 2007/2008 and tested for RSV by polymerase chain reaction. Of 1002 samples, only 4 tested positive (2 from a patient, 2 from staff). Sequence analyses of the G protein demonstrated that nosocomial transmission did not occurr between these individuals.