Transfusion-associated viruses have been, and will remain a major public health problem. Massive research efforts have reduced the threat to blood product recipients through the discovery of a number of etiologic agents, and the subsequent development of specific serologic tests for these viruses. Hepatitis C, hepatitis B, and human immunodeficiency virus (HIV) all pose a much smaller threat because of these advances. Although markedly lessened, the risk of transfusion-associated infection from both well-known agents and from emerging pathogens is still a concern for the general public. Exquisitely sensitive nucleic acid detection assays have been developed in the last decade but have not become a part of a routine screening protocol for protecting the nation's blood supply. This research plan proposes a straight-forward strategy for pooling serum specimens from large asymptomatic populations in order to reliably and unequivocally identify infectious donors. A two-tiered algorithm for screening blood products is outlined in the first section of this proposal. In the first tier, serum specimens are pooled and the resulting pools tested with a PCR-based assay. In the second tier, pools testing positive would then be broken down, and every individual donor from that pool would then be tested individually. A mathematical model based on this approach is presented, and serves as a useful guide for designing pooling strategies based on the expected prevalence of a disease within a population of interest. The experimental goals of this project are to develop protocols that allow the fractionation and concentration of transfusion associated viruses. A second goal is to design stabilizing formulas for viral nucleic acids to allow easy transport of specimen to reference laboratories. A third goal is to develop nucleic acid standards to control for falsely negative tests. The ultimate objective is to manufacture a kit for regional blood banks that contains an internal RNA/DNA control, a viral concentration solution, and a nucleic acid transport solution. PROPOSED COMMERCIAL APPLICATION: The goal of this research is to develop FDA-approved commercial kits that blood banks use to concentrate and transport serum samples for PCR testing. A potential, although smaller market for this technology may be present in the research Community.