Aedes aegypti L. is considered to have a proclivity for feeding on human blood even when other hosts are available. However, few studies have demonstrated host use by this mosquito in the continental United States, where local transmission of dengue, Zika, and chikungunya viruses has been recently documented. This study investigated the bloodmeal sources of female Ae. aegypti in the subtropical city of Key West and the surrounding county in Florida with the goal of identifying preferred hosts. Blood-engorged Ae. aegypti mosquitoes were collected from BG Sentinel traps used as part of a routine surveillance program in the Florida Keys (Monroe County, Florida). Bloodmeal samples were analyzed using PCR assays, sequencing, and comparison with reference sequences in GenBank. Aedes aegypti females from Key West fed predominantly on humans (79.6%) and did not differ statistically from females collected from the rest of the Florida Keys (69.5%). Culex quinquefasciatus Say (Diptera: Culicidae), considered a host generalist, was collected and analyzed from the same sites for comparative purposes. Females of Cx. quinquefasciatus fed predominantly (70.7%) on birds and nonhuman mammals in the Florida Keys, corroborating the validity of molecular assay breadth and demonstrating that given the same group of available hosts Ae. aegypti selects humans. Our results indicated that Ae. aegypti has a high rate of human-biting in a subtropical area within the United States, supporting its role in recent local transmission of dengue and other viruses.