Introduction: Blood is a scarce, but life saving resource. At the same time, unsafe transfusion practice also puts millions of people at risk of Transfusion Transmissible Infections (TTIs). Aim: To determine the prevalence and trend of blood borne infection namely Human Immunodeficiency Virus (HIV) among blood donors donating blood in a tertiary care hospital of Haryana, India. Materials and Methods: A retrospective study was conducted from September 2012 to August 2014 to calculate the prevalence and trend of HIV positivity among blood donors over a period of five years (2008-2012). Results were expressed in terms of frequency and percentage. Results: There were a total of 1,59,077 donations, including both voluntary and replacement donors. The overall prevalence of HIV infection among blood donors was found to be 0.26% over a period of five years (2008-2012). The HIV prevalence among blood donors showed an increased trend from 2008 (0.18%) to 2011 (0.34%), and then there was a dip in the year 2012 (0.23%). Conclusion: The number of female blood donors were less in comparison to male blood donor. The maximum number of donors were voluntary males followed by replacement males.