Sepsis (a toxic condition caused by the spread of an infection into the blood or tissues) and community acquired pneumonia (CAP) are high prevalence conditions and major cause of mortality in the hospitalized patient population. Combined, the two conditions affect millions of patients per year in the United States, are responsible for over a hundred thousand deaths and accounts for tens of billions of dollars in annual health care costs. The proposed research program is directed towards the development of better diagnostic tools that, by providing earlier and more accurate detection, will lead to more effective therapy and the lowering of overall health care costs. ? ? The long-term objective of this research proposal is the development of a point-of-care (POC) diagnostic for measuring sepsis and CAP. High sensitivity and specificity will be provided through the multiplexed measurement of multiple different markers of sepsis and CAP, including both the direct measurement of the causative infectious agents as well the measurement of characteristic markers of the host immune response. The work will leverage prevoious and ongoing work at Meso Scale Diagnostics (MSD) towards the development of a small automated POC instrument that conducts rapid quantitiative multiplexed measurements on inexpensive single-use cartridges. ? ? The proposed project has four specific aims: (1) measuring by electrochemiluminescence immunoassay five antigenic markers of pathogenic organisms and 35-40 protein markers of host immune response in a number of CAP, sepsis and control patients to identify markers that are highly predictive of sepsis and CAP; (2) developing approaches for the rapid generation, screening and selection of monoclonal antibodies against multiple subtypes of a pathogen for use in the development of assays for directly measuring infectious agents involved in sepsis and CAP; (3) transfer and demonstration of selected immunoassays from aims 1 and 2 to MSD's POC diagnostic system and (4) demonstration of multiplexed PCR assays on MSD's POC diagnostic system. The system will be expandable to allow introduction of additional markers in the future. Completion of the project will result in a working assay system that can be taken into clinical trials. ? ? ?