Zika virus (ZIKV) is an RNA flavivirus (Flaviviridaefamily) endemic in tropical and subtropical regions that is transmitted to humans byAedes(Stegomyia) species mosquitoes. The two main urban vectors of ZIKV areAedes aegyptiandAedes albopictus,which can be found throughout Brazil. This study investigated ZIKV infection in mosquito species sampled from urban forest fragments in Manaus (Brazilian Amazon). A total of 905 non-engorged femaleAe. aegypti(22 specimens) andAe. albopictus(883 specimens) were collected using BG-Sentinel traps, entomological hand nets, and Prokopack aspirators during the rainy and dry seasons between 2018 and 2021. All pools were macerated and used to inoculate C6/36 culture cells. Overall, 3/20 (15%)Ae. aegyptiand 5/241 (2%)Ae. albopictuspools screened using RT-qPCR were positive for ZIKV. No supernatants fromAe. aegyptiwere positive for ZIKV (0%), and 15 out of 241 (6.2%)Ae. albopictuspools were positive. In this study, we provide the first-ever evidence ofAe. albopictusnaturally infected with ZIKV in the Amazon region.