Abstract Objective: To determine the ability of early antigen detection, isolation of infected infants with Respiratory Syncytial Virus (RSV) and applied of standard infection control procedures to prevent nosocomial spread of virus from isolation rooms to the Neonatal Intensive Care Unit (NICU) where high-risk infants were under care. Methods: The study was performed between 1 October 2010 and 31 March 2011, prospectively. All newborns who were admitted to the NICU for Lower Respiratory Tract Infection (LRTI) (Group 1) were first tested using anusing Respi-Strip antigen detection kits to identify RSV (+) neonates. RSV (+) neonates with LRTI (Group 1a) were isolated and segregated in a room separate from RSV (-) neonates with LRTI (Group 1b). Infants at high risk for RSV infection who had been hospitalized continuously since birth (Group 2) were also segregated and observed at least weekly as sentinels for nosocomial spread of RSV infection. Standard infection control procedures were in place throughout the study period. Findings: Forty-one newborns were hospitalized and treated with a diagnosis of LRTI during the study period. Fifteen (37%) of these were RSV antigen-positive. No RSV outbreak or infection was observed with isolation and standard infection control procedures in the risk group premature infants during their hospitalization. Also this study show that; RSV related LRTI is high in neonatal period (37%). Conclusion: This study showed that isolation of infants with RSV and strict adherence to infection control procedures might prevent nosocomial spread of RSV infection in the NICU that care for at risk infants even in the absence of palivizumab prophylaxis.