In the present study, we used a competition ELISA (cELISA) assay to estimate the seroprevalence of HBoV1-4 in healthy Chinese individuals ranging in age from 0 to 70 years in Beijing, China. We also compared the seroconversion of anti-HBoV IgG antibodies in 31 paired serum samples from ARTI children who were positive for HBoV1 by PCR. Our findings provide informative data for evaluating the prevalence and pathologic roles of HBoVs.