Background. The COVID-19 was first reported in 2019 to cause pneumonia in people of Wuhan, Hubei province, China, is now associated with high mortality worldwide. Phylogenetic analysis revealed that SARS-CoV-2 (2019-nCoV) is closely (88%–89% similarity) related to the coronavirus circulating in Rhinolophus (horseshoe bats). More than 50 bat species belonging to eight families have been reported from Iran of which five species belong to the Rhinolophidae family. So far, no study has been done on COVID-19 infection in Iranian bats. Aim. The current study was performed, for the first time, to investigate the infection of Iranian bats with SARS-CoV-2. Methods. This cross-sectional study was conducted in 2021 using 183 bat samples collected from three caves in the south (Fars province) and two caves in the northwest (Kermanshah and Kurdistan provinces) of Iran. Bats’ digestive and respiratory system samples were collected from each bat of different species. The samples were evaluated by real-time PCR and by targeting a 221 bp fragment of the envelop (E) genes of SARS-CoV-2. Results. COVID-19 was detected in alimentary specimens of two of the Mediterranean horseshoe (Rhinolophus Euryale) bats. Conclusion. Although, based on the findings of the molecular evaluation, the infection of bats with COVID-19 was determined in this study, further studies are needed on a larger number of bats, particularly horseshoe bats, to confirm the potential infection of Iranian bats with COVID-19.