Hepatitis B, Hepatitis C and HIV infections represent global health problems of significant magnitude. There are studies and hypothesis which suggest that genetic predispositions like ABO and Rh blood group may affect occurrence of these diseases. The aim of this study was to find the prevalence of ABO blood groups, seroprevalence of hepatitis B, hepatitis C, HIV, syphilis, and malaria infections in healthy blood donors in our center, and to determine any association between different blood groups and seroreactivity. Blood donors are ideal for this purpose, as all donors are routinely subjected to ABO blood group phenotyping and screening for all these infective markers.This was a retrospective cross sectional study conducted at Regional Blood Transfusion Center at Dr. BSA Hospital for the years 2016 and 2017. All the donor samples were mandatorily tested for HBsAg, HIV 1&2 Ag-Ab and 2 Ag-A band HCV Ag-Ab using fourth-generation kits. TPHA method was used to test for syphilis. Malaria was tested using rapid-pan malaria card. Blood group was determined by forward and reverse grouping of donor sample using gel card method. Results of total 21,876 donors registered in the year 2016 and 2017 were analyzed. Total 320 (1.462%) samples were found to be reactive for HBsAg, 62 (0.283%) samples were found reactive for HCV and 47 (0.215%) samples were found reactive for HIV. 0.027% samples tested reactive for syphilis. Group-wise seroreactivity was also analyzed and it was found that the highest percentage of HBsAg and HCV reactivity was found in O positive donors and highest percentage of HIV reactivity was found in A-positive donors. Lowest seroreactivity amongst Rh-positive donors was found in AB-positive donors. Amongst Rh-negative donors, highest seroreactivity for HBsAg and HCV was found in B-negative donors. O-negative, AB-negative and A-negative showed no seroreactivity for either HCV, HIV or syphilis infection. All donors tested negative for malaria.