Dengue virus is one of the emerging agents that can be transmitted via blood transfusion from infected blood donors to recipients. In Malaysia, the increase in dengue infection may contribute to the existence of asymptomatic blood donors and increase the risk of blood supply contaminated with this virus. The aims of this study were to investigate the prevalence of NS1 dengue antigen among blood donors and to ascertain the demographic data of blood donors in Penang and and Perak. Methods: A total of 374 voluntary blood donors were recruited from two blood donation campaigns organised by Hospital Pulau Pinang, Penang and Hospital Raja Permaisuri Bainun, Ipoh, Perak from April to May 2016. From each centre, 187 voluntary blood donors were enrolled, blood was collected and Dengue NS1 Ag was screened on all the samples using Platelia dengue antigen test kit from Bio-Rad Laboratories, France. Results: All 374 samples were found to be negative for the Dengue NS1 antigen. Demographic data of these blood donors showed that the most common blood group was O Rh positive, men donated more than women and Chinese blood donors were the biggest group of donors. Conclusion: Even though dengue is endemic in Malaysia, none of the blood donors was screened positive for dengue NS1 antigen in the areas studied. This indicates that none of the blood donor at the time of donation was in viraemia stage. The established donor screening program ensures that the dengue transmission through transfusion is minimal in the areas studied.