Air purifiers with high-efficiency particulate air (HEPA) filters remove not only particulate matter but also airborne microorganisms in indoor environments. We investigated the bacterial community in HEPA filters (used for 1 year) and that in the floor dust of 12 office rooms in Beijing. We found that the viable bacteria proportion in the filter was significantly higher than that in the floor dust (p < 0.001). The Non-Metric Multi-Dimensional Scaling analysis showed that the bacterial communities in the filters and dust were significantly different (p = 0.001). The Chao1, Shannon-Wiener and phylogenetic diversity values in the filter were significantly higher than those in the dust (p < 0.001). The predominant bacterial classes in the filter were Alphaproteobacteria and Actinobacteria, whereas those in the dust were Bacteroidia, Clostridia and Bacilli. Human occupancy contributed more to the bacterial community in the filter than that in the dust. Klebsiella and Alloprevotella in the dust and filters positively correlated with the occupancy density. Soil bacteria contributed to a significantly higher proportion of the bacteria in the HEPA filter (p < 0.001). In contrast, human oral, indoor air and outdoor haze contributed to a higher proportion of the bacteria in the dust samples (p < 0.001, p < 0.01 and p < 0.05, respectively). As HEPA filters serve as an ecological niche for indoor bacteria, they should be carefully investigated during the assessment of indoor environmental health.