Hepatitis B is an infectious disease of great public health importance. Nigeria is one of the countries with the highest incidence of Hepatitis B Virus (HBV) infection worldwide. However, the accessibility and affordability of HBV DNA quantification (viral load) assay which is the key laboratory test for therapy initiation, and monitoring is a challenge to HBV management. The study was done to evaluate serum levels of HBV-DNA concentration and HBsAg titers of hepatitis b virus-infected subjects at NAUTH Nnewi. Cross sectional study design was used with a total of 264 subjects comprising of 88 HBsAg seropositive treatment naïve subjects, 88 HBsAg seropositive subjects on antiviral therapy as case subjects and 88 age-matched apparently healthy HBsAg seronegative individuals were recruited as control subjects. Hepatitis BVirus DNA assay was performed using real time PCR technique, ELISA technique was used for Hepatitis B surface antigen quantification, Hepatitis Bcore Antibody Immunoglobulin M and Hepatitis D Virus Immunoglobulin G assay. Immunochromatography was used for HBV Panel, Hepatitis C Virus assay, Human Immunodeficiency Virus testing. HBsAg quantification showed strong positive correlation with HBV DNA viral load both in treatment and nontreatment groups (r = 0.673; p < 0.001). The non-treatment group has higher viral load (M=805.50IU/ml) compared with treatment group (M = 65.50IU/ml) (p<0.001).