Background: Worldwide, 5–10% of people with chronic hepatitis B virus infection are co-infected with hepatitis D virus. In Qatar, there are no data on hepatitis D virus infection among patients positive for hepatitis B surface antigen (HBsAg). Aims: To determine the seroprevalence of hepatitis D virus infection among patients with chronic hepatitis B virus infection in Qatar and assess the characteristics of these patients. Methods: This was retrospective cohort study of all HBsAg-positive individuals tested for hepatitis D virus between 1 January 2010 and 29 December 2019 within Hamad Medical Corporation. Data were retrieved from electronic records and included demographic and clinical information of the patients. Results: Of 2348 HBsAg-positive patients, 125 were positive for hepatitis D virus (seroprevalence 5.3%). The median age of hepatitis D positive patients was significantly higher than patients negative for hepatitis D virus (P = 0.001). Most patients with hepatitis D had a hepatitis B viral load < 2000 IU/mL (53.6%) and were negative for hepatitis B e antigen (93.6%). A significantly greater proportion of patients positive for hepatitis D virus infection than those negative were infected with hepatitis C virus (P < 0.001), and had liver cirrhosis (P < 0.001) and hepatocellular carcinoma (P = 0.006) Conclusions: Hepatitis D virus infection is associated with lower hepatitis B virus viraemia and more advanced liver disease.