Hepatitis C virus (HCV) is an important causative agent of chronic liver disease worldwide distributed. In Tunisia, reported HCV seroprevalence is about 0.7%, with higher infection rate in the North-East region (Béjà). Subtype 1b is the largely predominant genotype. As it was suggested in a previous study, a specific HCV variant, subtype 1b was circulating in Tunisia, especially in urban areas of the North-Nest region. The aim of this work was to assess phylogenetic relatedness between viruses circulating in different other parts of Tunisia, and to compare them with those from the North-West region, and with those from other countries. Phylogenetic analyses were carried out on two viral regions: the NS5B and the E1. Phylogenetic analyses identified a group of sequences forming a cluster including almost exclusively Tunisian strains. This phylogenetic cluster comprises more than the half of all investigated Tunisian strains, especially those from urban parts of Béjà (North-West). Such results were observed not only after investigations in the NS5B region, but also after analyses in the E1 viral region. All these observations confirm the hypothesis about the specific local variant of HCV subtype 1b in the country, particularly in the North-West. This local variant could be related to a common HCV transmission route, as it was suggested in a previous publication.