The distribution of hepatitis B virus genotypes in Saudi Arabia is largely unknown. To the best of our knowledge there are no data available about HBV genotypes in southwestern region of the country. This study aimed to determine the epidemiologic distribution of hepatitis B virus genotypes in chronic hepatitis B patients in southwestern region, and to verify possible correlations between these genotypes and the clinical symptoms. A total of 160 patients with chronic hepatitis B infection were enrolled in this study. Sera were tested for liver function tests, hepatitis B virus markers and DNA load by standard procedures. HBV genotyping was performed by 2-tube nested PCR for determination of six genotypes (A-F). Genotype D was the most common, found in 135 (84.4%) patients, followed by A (18; 11.3%) and E (7; 4.3%). The rate of HBeAg positivity in genotype D patients was significantly lower compared with that in genotype A and E patients (p=0.01). There was no significant association between HBV genotypes and age, gender, liver function tests, or HBV DNA load. Genotypes D and E were predominant in chronic hepatitis B patients in southwestern Saudi Arabia. Awareness of hepatitis B virus serologic and genotypic patterns might help in the formulation of management plans, predicting clinical outcomes and updating prevention strategies.