22 23 *gaspary.mwanyika@sacids.org 24 25 26 27 PLOS Neglected Tropical Diseases 2 28 Abstract 29 Background 30 Dengue is an important mosquito-borne viral disease in tropical and sub-tropical 31 countries. In this study molecular characterization was carried out to determine 32 dengue viruses circulating among patients at health facilities during 2019 epidemic in 33 Dar es Salaam, Tanzania. 34 Methods 35 The study involved outpatients seeking care for febrile illness at four health facilities 36 in Kinondoni and Ilala Districts of Dar es Salaam City in Tanzania. A total of 45 sera 37 from the outpatients were confirmed dengue-positive for dengue virus (DENV) non-38 structural protein 1 (NS1) antigen and/or NS1-IgG/IgM antibodies using on-site rapid 39 test. The presence of the virus was detected by reverse-transcription polymerase 40 chain reaction (RT-PCR) method. Of the 45 sera, 20 samples were selected 41 randomly for identification of specific dengue virus serotypes using RT-PCR followed 42 by evaluation of resulting amplicons on agarose gel electrophoresis. 43 Findings and significance 44 Both Dengue virus serotypes 1 (DENV-1) and 3 (DENV-3) were detected in the 45 samples tested with the former being dominant. We present the first evidence of 46 dengue virus co-infection of DENV-1 and DENV-3 serotypes in Tanzania. The 47 emergence of DENV-1 indicates the possibility of importation of the virus to Tanzania 48 from endemic countries. Due to DENV serotype co-circulation, there is an increased 49 risk of severe dengue in future epidemics. Our findings advocate the importance of 50 genomic-based surveillance to provide rapid evidence of dengue virus 51 emergence/re-emergence and spread. 52 Author Summary 54 Dengue viruses are the most important mosquito-borne pathogens that pose a 55 serious global health threat. Tanzania has reported several dengue virus epidemics 56 since 2010 with the majority of the epidemics occurring in Dar-es-Salaam city. Until 57 August 2019, a total of 6,859 dengue cases have been confirmed in the country. We 58 performed molecular characterization of dengue viruses (DENV) circulating during 59 the 2019 epidemic phase. It was found that DENV-1 serotype was dominant during 60 the epidemic and two samples of the tested sera were co-infected by DENV-1 and 61