Abstract Background Human infection with influenza A(H1N1) 2009 was first identified in the United States on 15 April 2009 and on 11 June 2009, WHO declared that the rapidly spreading swine-origin influenza virus constituted a global pandemic. We evaluated the seroprevalence of influenza A(H1N1) 2009 virus on a large public University campus, as well as disparities in demographic, symptomatic and vaccination characteristics of participants. Methods Using a cross-sectional study design, sera was collected from volunteers and then tested for the presence of antibodies to the virus using a ≥ 1:40 dilution cut-off by hemagglutination inhibition assay. In conjunction, participants were asked to complete a questionnaire allowing us to estimate risk factors for infection in this population, as well as distinguish artificially derived antibodies from naturally derived antibodies. Results 300 total participants were recruited and tested. 158 (52.6%) tested positive for influenza A(H1N1) 2009 via hemagglutination inhibition assay using a ≥ 1:40 dilution cut-off. 86 people (54.4%) tested positive for H1N1 but did not report experiencing symptoms during the pandemic meeting the May 2010 CDC definition of influenza-like illness. Furthermore, of those individuals who reported that they had received the H1N1 vaccine, 16% did not test positive. Conclusions Overall, 52.7% of the total study population tested positive for influenza A(H1N1) 2009. 54.4% of those who tested positive for influenza A(H1N1) 2009 using the ≥ 1:40 dilution cut-off on the hemagglutination inhibition assay in this study population did not report experiencing symptoms during the pandemic meeting the May 2010 CDC definition of influenza-like illness. 16% of those who reported receiving the H1N1 vaccine did not test positive by HAI. We also found that vaccination coverage for H1N1 vaccine was poor among Blacks and Latinos, despite the fact that vaccine was readily available at no cost.