Journal of Pathogens and global healthDengue is an important mosquito-borne viral disease in humans in tropical and subtropical countries. In 2019, a total of 6917 dengue cases were reported in Tanzania based on serological analysis. The aim of this study was to confirm the presence of dengue virus (DENV) and conduct its genetic characterization. A total of 191 serum samples were collected from the outpatients seeking care from health facilities in Kinondoni and Ilala districts between March and May 2019. All the samples were initially tested for the presence of non-structural protein 1 and anti-DENV immunoglobulin G (IgG) and IgM using a commercial OnSite Duo Dengue Ag-IgG/IgM rapid test. Of the 191 sera, 110 (57.6%) were DENV seropositive. The presence of DENV ribonucleic acid was confirmed in 18.2% of the seropositive sera by reverse transcription polymerase chain reaction (RT-PCR). The RT-PCR products were cleaned and partial sequences of DENV poly­ protein gene determined using dideoxynucleotide cycle sequencing followed by phylogenetic analysis. We present the occurrence of DENV serotype 1 (DENV-1) during the 2019 outbreak in Tanzania. The DENV-1 strains reported in the present study are highly identical and cluster with Asian DENV-1 strains indicating the possibility of intercontinental spread of DENV through globalization. We advocate for the need for molecular surveillance of dengue viruses during outbreaks to provide rapid evidence of the disease to guide public health interventions.