Hepatitis B Virus genome evolves at an error rate of ~ 10 –3 to 10 –6 nucleotide substitutions/site/year, which is approximately 100 times higher than that of other DNA viruses. Accumulation of these mutations over a long period of time results in a large amount of genetic diversity. It is important to note that the rates of disease progression, clinical and treatment outcomes may different for each HBV genotype. Our aim was to detect the most prevalent genotypes of Hepatitis B virus among patients with liver disease in KD Medical College Mathura. A total of 280 patients with liver disease who were also positive for HBsAg by Elisa were enrolled. All sera were tested for liver function test, HBe Ag and HBV DNA viral load. HBV genotyping was done by RT-PCR. The median of HBV DNA viral load and ALT levels were 4.56 log IU/mL and 106.95 IU/L, respectively. HBV genotype D (38.18%) was the predominant circulating genotype, followed by genotypes A (21.81%), B (3.6%), C (0.9%), and Genotypes A/D (7.27%), D/C (4.54%), B/D (5.45%) and A/B (2.7%) were present. This study illustrated distribution of HBV genotype A, B, C, D. HBV in Mathura is comprised of a mixture of HBV genotypes. Genotyping can help practicing physicians identify those at risk of disease progression and determine optimal anti viral therapy. The study contributes in narrowing the existing gap of HBV molecular study in Uttar Pradesh India