Chikungunya virus (CHIKV) is a mosquito-borne alphavirus transmitted by Aedes aegypti and Aedes albopictus. Despite favorable ecological conditions in southern Iran, data on CHIKV exposure in Bushehr Province are lacking. This study aimed to determine the seroprevalence of CHIKV-specific IgG antibodies and assess potential viral circulation. A cross-sectional seroepidemiological survey was conducted from July to September 2023 in ten cities across Bushehr Province. Serum samples (n = 180) collected from volunteers at hospitals and blood donation centers were tested for CHIKV-specific IgG antibodies using a commercial ELISA kit. Of the 180 serum samples analyzed, five tested positive for CHIKV IgG antibodies, yielding an overall seroprevalence rate of 2.78%. Seropositive individuals were identified in Bushehr, Dashtestan, and Dashti cities, indicating localized and possibly silent transmission. This study provides the first serological evidence of CHIKV exposure in Bushehr Province. The findings underscore the potential for local transmission in areas where competent Aedes vectors are present and highlight the urgent need for sustained vector surveillance, early warning systems, and public health preparedness in southern Iran.