Drug resistance to Human Immunodeficiency Virus (HIV) is a major factor in treatment failure. Although this resistance is mostly related to drug exposure, treatment-naïve patients living with HIV have also been shown to develop primary drug resistance. Resistance monitoring before treatment prevents the first-line antiretroviral therapy (ART) failure and improves treatment outcomes. Very few data are available on the prevalence of HIV drug-resistance in Tunisia, especially among ART-naïve patients. This study aimed to investigate HIV-1 drug resistance patterns among ART-naïve Tunisian and African individuals. The study cohort included 77 treatment-naïve PLHIV who visited the HIV Center of Monastir to initiate ART. Viral RNAs were partially sequenced in the RT and PR regions using Sanger sequencing. Subtypes and Drug Resistance mutations (DRMs) were determined using the Stanford HIV Database. HIV-1 subtying revealed the predominance of the CRF02_AG, followed by the subtype B. 12,9% of variants were shown to have at least one Surveillance DRM (SDRM) in the PR/RT region. Most observed resistance cases (60%) were noted for RT inhibitors, with K103N the most involved SDRM-inducing resistance to first and second generation NNRTs (efavirenz, nevirapine, and rilpivirine). In the PR gene, resistance was found in four samples on 46 and 85 codons, affecting the sensitivity for Atazanavir and Lopinavir drugs. This study provides the first baseline information for pre-treatment drug resistance surveillance of HIV-1 variants in Tunisia, and shows considerable mutations, which may impede patient management.