The potential introduction of SARS-CoV-2, the virus responsible for the COVID-19 pandemic, into North American bat populations is of interest to wildlife managers due to recent declines of several species. Populations of little brown bats (Myotis lucifugus) have collapsed due to white-nose syndrome (WNS), a disease caused by the introduction and spread of the fungal pathogen Pseudogymnoascus destructans (Pd). Throughout much of the United States and southern Canada, large colonies of the species routinely established diurnal roosts in anthropogenic structures creating the potential for direct human contact and cross-species disease transmission. Given recent declines and the potential for further disease impacts, we collected oral swabs from eight little brown bat colonies for RT-qPCR analysis to describe the presence and prevalence of SARS-CoV-2. We visited colonies in Maryland (n = 1), New Hampshire (n = 1), New Jersey (n = 2), New York (n = 1), Rhode Island (n = 2), and Virginia (n = 1) during May-August, 2022. We have analyzed samples from 235 individuals and all have tested negative for SARS-CoV-2. Our results indicate that little brown bats are either negative for SARS-CoV-2 or that it persists in undetectable levels in populations of the Mid-Atlantic and Northeast in the summer months. Nonetheless, future work addressing other seasons or using serologic approaches may still be warranted to conclusively determine disease status.