Abstract: Objective To investigate the prevalence of common respiratory viruses in infants and young children with acute respiratory infection (ARI) during the pandemic of 2009 influenza A(H1N1) in Beijing． Methods Primers and probes were designed to develop a novel multiplex real-time PCR for the detection of 7 respiratory viruses including respiratory syncytial virus (RSV) types A and B, parainfluenza virus (PIV) types 1, 2 and 3, adenovirus (ADV) and human bocavirus(HBoV) in clinical samples. Clinical specimens were collected from children who visited the Children′s Hospital affiliated to Capital Institute of Pediatrics with ARI from June 2009 to February 2010 and had been tested negative for 2009 influenza A(H1N1). Results The method could detect as few as 10-300 copies of each target plasmid DNA, and could only specifically detect the target virus but not other viruses in specimens. The application of the method in 849 throat swab specimens indicated that the overall positive rate was 39.0%, 1.4% for RSV-A, 8.4% for RSV-B, 8.2% for PIV-1, 0.5% for PIV-2, 3.9% for PIV-3, 13.9% for ADV, 2.7% for HBoV, respectively. In the tested positive cases, 87.6% cases were under 5 years of age. Statistical analysis of the data demonstrated that the dominated subtype of RSV was RSV-B (85.5%) which caused an epidemic peak from Nov. 2009 to Feb. 2010; PIV infections were mainly associated with PIV-1 from Jul. 2009 to Oct. 2009 and with PIV-3 from Jun. 2009 to Sep. 2009; ADV had a relatively high detectable rate during the whole period (an average of 13.9%); and the detectable rate of HBoV increased from Sep. 2009 to Dec. 2009. Conclusions In addition to influenza virus, ADV, RSV-B and PIV infections were also associated with ARI in children during 2009 pandemic influenza A（H1N1）in Beijing. No evidence showed that the characteristics of both the seasonality and the morbidity of the viruses involving in this study were impacted by the pandemic of the new influenza virus H1N1.