To determine changes (Trends) in infection rates of Hepatitis B surface antigen (HBsAg), Hepatitis C virus (HCV) and Human immunodeficiency virus (HIV) in blood donors of Khulna Population. Specimens of 34953 voluntary blood donors & party or relative donors in Transfusion Medicine Department of Khulna Medical College Hospital from 2007 to 2009 were screened for HBsAg, anti-HCV, anti-HIV 1 & 2 reactivity in a cross-sectional survey by rapid test method. Reactive samples were verified using a recognized confirmatory test which consisted of a second generation enzyme immune assay (HBsAg), anti-HCV antibodies by anti-HCV EIA & for HIV by western Blot, respectively. The seroprevalence of HBsAg, Anti-HCV, HIV antibody 1 & 2 was 1.4%, 0.09% & 0.03% respectively in all blood donors. Prevalence of confirmed positivity was 0.62% for HBsAg, 0.04 % for Anti-HCV, 0.02% for HIV Western Blot. Between 2007 to 2009 a decreasing trend was observed in HBsAg frequency, HCV frequency decreased in 2009 compared to 2007. One HIV positivity found in 2009. Although the frequency of transfusion transmitted infections is low, party or relative donors have some risk factors than voluntary blood donors. Through more scrutiny in donor selection, improved serological test & reevaluation of infections routes in donor, infection reduction can be achieved.