Recent studies have suggested that microbial aerosolization in wildfire smoke is an understudied source of microbes to the atmosphere. Wildfire smoke can travel thousands of kilometers from its source with the potential to facilitate the transport of microbes, including microbes that can have far‐reaching impacts on human or ecosystem health. However, the relevance of longer‐range detection of microbes in smoke plumes remains undetermined, as previous studies have mainly focused on analyses of bioaerosols collected adjacent to or directly above wildfires. Therefore, we investigated whether wildfire smoke estimated to originate >30 km from different wildfire sources would contain detectable levels of bacterial and fungal DNA at ground level, hypothesizing that smoke‐impacted air would harbor greater amounts and a distinct composition of microbes as compared to ambient air. We used cultivation‐independent approaches to analyze 150 filters collected over time from three sampling locations in the western United States, of which 34 filters were determined to capture wildfire smoke events. Contrary to our hypothesis, smoke‐impacted samples harbored lower amounts of microbial DNA. Likewise, there was a limited signal in the composition of the microbial assemblages detected in smoke‐affected samples as compared to ambient air, but we did find that changes in humidity were associated with temporal variation in the composition of the bacterial and fungal bioaerosols. With our study design, we were unable to detect a robust and distinct microbial signal in ground‐level smoke originating from distant wildfires.