Abstract: Hepatitis B virus infection has become a major health problem world over. The present study was designed to investigate the prevalence of HBV-markers in relation to promotive factors among human urban population of Bahawalpur-district, Pakistan. Randomly selected population (both sex, 10-70 years) of Bahawalpur (n = 1464) divided into 3 age groups; young (10≥20 years), mature (20≥50 years) and old (>50 years) was tested for blood groups. HBsAg and/or Anti-HBc positive participants were interviewed regarding past and present life style to determine promotive factors. Over all HBV-markers prevalence was observed 12.57%. Maximum infection was 13.79% (95% CI: 11.09 to 17.03, p>0.01) in old group compared with mature and young groups. HBV-markers positivity was statistically not associated with sex, ABO blood groups and Rh-factors. An inverse relationship (p>0.01) was observed between prevalence of HBV-markers and socioeconomic level of under trial population. Significantly (p>0.01) high seroprevalence was found in individuals with high rate of crowding (>4/room), stored water users, regular parlor/barber’s shop visitors, those who were living/working with HBV infected people (s), surgically operated/blood transfused participants, dental clinic visitors and due to non-sterile pricking. HBV-infection is spreading rapidly among population of Bahawalpur district, Pakistan linking with deprived socioeconomic conditions, poor sanitation, non-sterile pricking, unprotected surgery/blood transfusion, parlor/barber’s shops, dentist’s clinics and HBV infected/carriers.