The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase I project is to enable and support the development, approval, and clinical use of new therapeutics the cure HIV-1 infection. HIV-1 infection can be controlled by available antiretroviral therapy, but cannot be cured due to the persistence of the virus in a quiescent, or latent state. This latent HIV-1 is the barrier to curing infection. While latent HIV-1 can be found in all infected individuals, it is present in very low frequencies. To cure HIV-1 infection, new therapeutics that eliminate latent HIV-1 must be developed. This development is hindered by current lack of a sensitive, accurate, and scalable test to measure latent HIV-1 in infected individuals. The propose project seeks to develop and optimize a sensitive, accurate, and scalable test to measure latent HIV-1 in the blood of infected individuals. Current tests used in research laboratories to measure latent HIV-1 are either non-specific or require large volumes of blood and many days to complete. This project is focused on developing a new molecular test that specifically measures latent HIV-1 from a minimal volume of blood in hours. Specifically, the project is focused on optimizing the preparation of blood samples and the reaction conditions necessary for robust performance of this new molecular test for latent HIV-1, as well as collecting samples from HIV-1 infected individuals to support future validation studies. Successful completion of this project will yield a latent HIV-1 test prototype that can be rapidly validated and advanced towards eventual clinical use.