In alignment with Morocco's national strategy for eliminating viral hepatitis, we aimed to characterize and update the virological profile of chronic hepatitis B patients. Demographic, serological and molecular parameters of 804 HBsAg-positive patients were retrospectively analyzed. Overall, 58.24 % were HBV-positive (55.37 % males, p = 0.74). The median age was 46 years (37-57). Patients ≤ 24 years comprised 5 % of HBsAg-positive and 4.34 % of HBV-positive cases. The median viral load was 2.62 log 10 IU/mL (1.87-3.44). The prevalent genotypes were D (91.04 %), A (7.55 %) and E (1.41 %). Liver enzymes were normal in most of cases. 91.04 % of patients were HBeAg-negative, with 92.23 % having genotype D (p < 0.001). Co-infection rates with other hepatitis viruses were low. Significant associations were found between HBeAg-negative status, genotype D, viral load, and liver enzyme levels (p < 0.001). We highlighted the need for prenatal HBsAg screening for pregnant women and prioritizing the birth-dose vaccine to prevent mother-to-child transmission, especially after the COVID-19 pandemic.