Awards issued prior to january 20, 2025, were funded under previous administrations and may not reflect the priorities and policies of the current administration.** microbial foodborne illness sickens…

Awards issued prior to january 20, 2025, were funded under previous administrations and may not reflect the priorities and policies of the current administration.** microbial foodborne illness sickens millions and kills thousands in the U.S. Annually. One of the major challenges to preventing the transmission of foodborne pathogens is the lack of ideal rapid, portable, and sensitive detection of pathogens from foods samples. Ligand-based detection techniques are portable, rapid but lack the degree of sensitivity needed for detection of pathogens in foods. Nucleic acid amplification-based techniques are highly sensitive, but lack portability and are sensitive to sample-associated inhibitors, thus requiring extensive sample purification. Isothermal amplification techniques like lamp and RPA show promise because of advantages, one of which is their relative tolerance of inhibitors, allowing for crude extraction/purification in-field. However, these methods can sometimes lack quantitative ability. Similarly, digital polymerase chain reaction (dpcr) also has exhibited tolerance for inhibitors and better quantitative ability, but lacks portability. In this proposal, we seek to develop and evaluate a nanoliter reactor that both digitalizes and detects any nucleic acid amplification-based technique in a portable format. We will explore the degree to which digitalization of RPA and lamp chemistries in a fully automated, in situ chip can aid rapid detection with the need for only crude sample extraction.