**5. Conclusions**

Despite the sample size being a limitation of the study, this suggests that the spread of HCV1b was maintained in the Calabria Region by sporadic infections, mainly acquired through the unsafe use of glass syringes, dental treatment and surgical intervention. Even if our analysis was performed on samples collected in 2015–2016, the frequency of natural RASs carried on subtype-specific viral strains, as well as comorbidities of treated patients, should be taken into account for the effectiveness of IFN-free regimens to eradicate HCV infection.

**Supplementary Materials:** The following are available online at https://www.mdpi.com/article/10 .3390/jcm10081655/s1, Figure S1: Likelihood mapping of HCV NS5B first (a) and second (b) dataset.

**Author Contributions:** A.R.C. and R.B.: conceptualization. A.C., S.T., M.E., A.L.P., G.P., G.S.B., A.Q., G.M.: methodology. A.C., S.T., M.E., A.L.P., G.P., N.M.: A.C., S.T., M.E., G.P.: formal analysis. investigation. E.M.T., C.C., M.M., F.S., C.T.: data curation. A.C., S.T., A.L.P., N.M.: writing—original draft preparation. A.R.C., M.C.L., C.T., N.M.: writing—review and editing. A.R.C.: supervision. All authors have read and agreed to the published version of the manuscript.

**Funding:** This research received no external funding.

**Institutional Review Board Statement:** The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Ethical Committee (#100; 27 April 2017) of the "Mater Domini" University Teaching Hospital of Catanzaro, Italy.

**Informed Consent Statement:** Patient consent was waived due to non-interventional study. The Ethical Committee approved the criteria that there is no need for informed consent for a noninterventional study.

**Data Availability Statement:** The 41 NS5B and 22 NS5A newly generated sequences used for molecular analysis have been uploaded to GenBank under the following accession numbers: MW357752- MW357814. Materials supporting the findings of this study are available within the article.

**Acknowledgments:** The authors would like to thank Neill J. Adams (Department of Health Sciences, Institute of Microbiology, "Magna Grecia" University of Catanzaro, Italy) who checked the English throughout the manuscript. Maria Mazzitelli was supported as PhD student by European Commission (FESR FSE 2014-2020) and by Calabria Region (Italy). European Commission and Calabria Region cannot be held responsible for any use, which may be made of information contained therein.

**Conflicts of Interest:** The authors declare no conflict of interest.
