The maintenance of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) among wildlife populations poses a potential risk for the emergence of novel variants. Therefore, monitoring SARS-CoV-2 infection among animals is crucial. As urban rodents live in close proximity to human habitats, there is concern that they may be a potential source of zoonoses. To examine the prevalence of SARS-CoV-2 in rodent populations, we analyzed 128 serum samples and 129 oral swabs collected from 128 brown rats (Rattus norvegicus) and 2 black rats (Rattus rattus) captured for pest control purposes in Tokyo, Japan, between May and December 2023. A virus-neutralizing test using the Omicron variant revealed no evidence of SARS-CoV-2 infection in these populations. Real-time RT-PCR from oral swabs did not detect any SARS-CoV-2 RNA-positive rats. These results indicate the low probability of SARS-CoV-2 circulation among rat populations in Tokyo.