Arboviruses transmitted by Aedes aegypti cause high number of cases and deaths annually. The aim was to investigate the presence of the presence of Dengue (DENV), Zika (ZIKV) and Chikungunya (CHIKV) viruses in endemic areas of Maranhão, northeastern Brazil. The study was carried out in Caxias, Codó, Peritoró, and São Mateus do Maranhão in 2019 (Caxias) and 2022. The blood samples were subjected to RNA extraction and then tested by RT-qPCR. Cell culture was used to attempt viral isolation and subsequent sequencing. In total, 171 samples were analyzed (32 from 2019, 18.7%) and 72 (42.1%) were found to have arboviruses: 68 (39.7%) from Caxias; 2 (1.1%) from Codó; 1 (0.6%) from Peritoró; and 1 (0.6%) from São Mateus. Overall, 85.3% (n = 58) of the positive samples were infected with DENV-1, 4 (four) (5.9%) with DENV-2 (Caxias), 1 (one) (1.5%) with DENV-3 (Caxias), and in 6 (six) (7.3%) samples CHIKV was detected, with one co-infection of DENV-1 and CHIKV (Caxias). The DENV-1 genotype V and the ECSA genotype of CHIKV were characterized in samples from Caxias. The detection of DENV-1, DENV-2, DENV-3, and more CHIKV in the interior of Maranhão alerts to the importance of virological studies in these areas.