Hepatitis B virus (HBV) infection is an important health problem and the major cause of chronic hepatitis, cirrhosis, and hepatocellular carcinoma (HCC) especially in Upper Egypt. The aim of this study was to estimate the prevalence of HBV among Assiut governorate patients. Our study examined 1085 patients for HBsAg at Assiut governorate, Egypt. Out of the 1085 population tested for HBsAg, 623 (57.5%) were males whereas 462 (42.5%) were females. A total of 165 out 1085 were seropositive for HBsAg (15.2%). The highest seropositive of HBsAg were recorded in 106/623 males (17.1%) compared as 59/462 females (12.8%). HBsAg seropositive were decreased with grassing age, the highest seroprevalence of HBsAg 62/165 (37.6%) was recorded in age ranges between 21 and 30 years, while the lowest seroprevalence of HBsAg 8/165 (4.8%) was recorded in age >60 year. All patients were divided into four groups according to HBV-DNA. Seroprevalence of HBsAg was increased with group 2 (Low titer of HBV-DNA <2000 IU/mL) (46.1%), where highest prevalence of HBsAg was recorded in males (37.6%) compared to as females (8.5%). The lowest seroprevalence of HBsAg were recoded with group 3 (high titer of HBV-DNA > 2000 IU/mL) (14.5%). HBV was common in rural versus urban community areas (78.2% versus 21.8%, respectively). Screening of HBV infection to monitor liver disease progression in HBV carriers by using molecular, biochemical, and serological markers, so that effective treatment can be initiated early before the development of advanced liver disease.