Abstract The lack of information on the occurrence of species in a region limits the understanding of the composition and structure of the local community and, consequently, restricts the proposition of effective measures for species conservation. In this study, we researched the reptiles in the Duas Bocas Biological Reserve (DBBR), Espírito Santo, southeastern Brazil. We analyzed the parameters of the local community, such as richness, composition, and abundance of species. We conducted samplings from August 2017 to January 2019, through active search. We performed the samplings in nine standard plots of 250 meters in length. All individuals located in the plots or occasionally on the trails were registered. To evaluate sample effort to characterize the reptile community, we performed an accumulation curve of species, and to update the DBBR reptile list, we used as secondary data specimens deposited in collections and previously published studies. Considering primary and secondary data, we recorded 38 species, one chelonia, 13 lizards, and 24 snakes. Our study showed a richness of 15 additional species to the previous list of reptiles, increasing by about 40% the known richness to the area. We concluded that the DBBR holds a high richness of reptile species, representing 12% of the richness of reptiles of the Atlantic Forest and with a considerable number of endemic species in this biome. This suggests that the DBBR is an important forest fragment constituting a reservoir of the biodiversity of the reptiles of the Atlantic Forest.