Background: Immunodeficiency Virus (HIV) and Hepatitis C Virus (HCV) share the same routes of transmission, therefore, co-infection by both viruses represents a challenge to the goal of eliminating viral hepatitis as a public health threat. There are an estimated 2.3 million people living with HIV/HCV worldwide. Most of these cases affect vulnerable populations located in places with low infrastructure. Because of this, the use of alternative samples such as Dried Blood on Spot (DBS) would facilitate access to diagnosis and HCV treatment. The aim of this study is to evaluate the HCV genetic variability in HIV/HCV individuals by correlating paired serum and DBS samples. Methods: A total of 14 HIV/HCV individuals, recruited from reference outpatient clinics in the city of Rio de Janeiro/Brazil, were included. From them, 64 % were man, mean of age 54±7. HCV RNA from both serum and DBS samples was RT-PCR amplified and sequenced with HCV NS5B-specific oligonucleotides. All positive samples were submitted to phylogenetic analysis. Results: Serum mean HCV load was 6.2 ± 0.5 log IU/mL. All patients presented undetectable HIV RNA. The distribution of HCV genotypes/subgenotypes was 1a (4/14); 1b (5/14); 3a (4/14); and 4d (1/14). Most paired serum and DBS samples showed concordant results (genetic distance: 0.0 to 0.16). One individual showed discordance in the subtypes between serum and DBS. Three individuals presented the 316 N Resistance Associated Mutation (RAS) in both serum and DBS. Conclusion: Our results demonstrate the applicability of DBS for HCV molecular tracking in HIV/HCV coinfected patients for viral genomic surveillance in key and vulnerable populations.