Objective: To determine frequency and trends of hepatitis B & C and Human Immune deficiency Virus (HIV) in blood donors presenting to the blood bank of a tertiary hospital. Study Design: Cross Sectional Descriptive study. Setting: Blood Bank of Khyber Teaching Hospital, Peshawar. Period: January 2014 to October 2019. Material & Methods: A total of 119263 blood bags were screened during the study period. Donors of age 18-55 years age and weight above 55 kilograms were included in the study. Cases who were drug addicts, anemic and jaundiced were excluded from the study. All blood bags were screened for Hepatitis B & C and HIV using automated ELISA Cobas E-114 equipment. The data was analysed and results were calculated in terms of percentages. Results: A total of 119263 blood donors presented to the blood bank during the study period. Age range of the study sample was 20-45 years with mean of 31±7 years. There were 119193 (99.94%) males and 70 (0.06%) females. The overall frequency of hepatitis B, hepatitis C and HIV was 1.42%, 0.76% and 0.018% respectively. The frequency of these infections was lowest in 2015 and highest in 2016.The trend of hepatitis B increased while that of hepatitis C decreased from 2014 to 2019. Conclusion: Hepatitis B infection is more prevalent in blood donors as compared to hepatitis C and HIV in our region. Actions should be taken to create awareness in population about the spread of hepatitis B and strict screening of the blood donors should be ensured to stop the rise in incidence of hepatitis B in our population.