Study disposition and enrolments. Total 288 consecutive HBsAg positive patient's samples were received at Genome Centre for Molecular based Diagnostics & Research. Of these total 98 patients samples were excluded from the study either the volume of sera were not sufficient for testing (n = 19) or failed to meet inclusion criteria of the study (n = 79) as they were HBV DNA negative by real-time PCR. Total 190 patients with chronic HBV who fulfilled the study criteria were enrolled for this study Out of 190 enrolled patients, 63.7% (n = 121) were males and 36.3% (n = 69) were females. Total 58(28%) samples were found positive with double infection of HBV &HDV. In the provinces of Sindh, Khyber Pakhtoonkhaw (KPK) and Punjab the observed prevalence of HDV was 67%, 6% and 4% respectively.