Introduction: Occurrence of detectable amounts of viral antigen or viral particles in the blood of infected patients with Hepatitis B virus (HBV) is a significant characteristic of HBV infection. Detection of HBV antigen or its DNA among individuals of a community is a crucial factor to know the burden of HBV infection. Hepatitis B surface antigen (HBsAg) is a suitable marker of HBV infection but a poor indicator of infectivity since its presence is not a direct measure of the presence of viable virions. Hence the tests for the detection of HBV DNA or HBsAg are used. The measurement of HBV DNA in serum has become the main tool to identify viral load, to monitor patients’ therapy and to predict whether antiviral therapy would be successful or not. Methods: The present study was designated to identify HBV infected individuals among adolescent age group by using combined methods namely, rapid immunoassay technique for HBsAg detection (HEPACARD) and Conventional Polymerase Chain reaction (PCR) analysis for HBV DNA detection. Results: Serum samples from 39 patients suspected of HBV infection were tested for the presence of HBsAg and HBV DNA. Eight specimens (20%) were positive for HBsAg as well as HBV DNA using PCR reaction. The ratio of spectrophotometric analysis of the extracted DNA samples was between 1.80-1.92 indicating a highly purified DNA. The gel electrophoresis of amplified PCR products of HBV DNA revealed a single 524 bp band in the test samples. Conclusion: Screening for HBV infection among adolescents by HEPACARD and further confirmation by PCR is recommended to monitor the progression of the disease and antiviral treatment.