Hepatitis B virus (HBV)-related Hepatocellular carcinoma (HCC) is a prominent form of cancer that ranks among the top malignancies globally, contributing significantly to cancer-related mortality rates (Villanueva, 2019). In HBV associated-HCC, the Hepatitis B Virus X protein (HBx) was reported to be a key regulator. It is a small multifunctional non-structural protein that plays a role in the biology and development of HBV by promoting virus replication and modulating the expression of host genes (Guerrieri et al., 2017). Although HBx was previously deemed irrelevant to the life cycle of HBV, it has since been recognized that it possesses transformative characteristics, thereby facilitating tumorigenesis. This can be attributed to its ability to enhance HBV replication, such as through HBx-dependent covalently closed circular DNA (cccDNA) formation or the modification of histones associated with cccDNA (Schollmeier et al., 2023). In addition, HBx plays a crucial role in the pathophysiology of HBV-related HCC (Sekiba et al., 2022). Indeed, an elevated risk of developing HCC is typically linked to mutations and deletions in the HBV genome. Of them, mutations in the precore-core (preC-C) regions are the most reported and are correlated with HBV genotype D, being the most common in the Mediterranean area, Morocco included, all along with the genotype A (Madihi et al., 2020a). However, the commercially available diagnosis assays for HBV in the same area target a sequence in the HBV genome that (preC-C) is associated with a large number of mutants related to the predominant genotype. Thus, these sheds light implicitly on a non-accurate diagnosis and raises the need for a more precise and specific diagnosis for such populations. The aim of this study consists to develop a Quantitative Real-Time Polymerase Chain Reaction Assay (qRT-PCR) for HBV DNA more adapted for a population with such characteristics. Finally, this study and after concretizing it, the African and non-African countries of interest could benefit from such assay.