** AWARDS ISSUED PRIOR TO JANUARY 20, 2025, WERE FUNDED UNDER PREVIOUS ADMINISTRATIONS AND MAY NOT REFLECT THE PRIORITIES AND POLICIES OF THE CURRENT ADMINISTRATION.** AFRICAN SWINE FEVER (ASF) IS A HIGHLY CONTAGIOUS AND OFTEN LETHAL INFECTIOUS DISEASE OF SWINE. CURRENTLY, STRICT CONTROL STRATEGIES REMAIN THE ONLY EFFECTIVE WAY OF ASF CONTAINMENT, TYPICALLY REPLYING ON EARLY DISEASE DETECTION THROUGH RAPID FIELD SUSPICION AND LABORATORY DIAGNOSIS FOLLOWED BY SLAUGHTERING ALL AFFECTED OR EXPOSED SWINE HERDS.THEREFORE, ASF HAS SERIOUS SOCIOECONOMIC IMPACT, ESPECIALLY IN COUNTRIES THAT EXPORT LIVE PIGS, PORK AND/OR PRODUCTS, AS WELL AS IN COUNTRIES WHERE THESE PRODUCTS ARE IMPORTANT SOURCES OF DIET. TO DATE, ASF HAS ALREADY SPREAD FROM SUB-SAHARAN AFRICA TO MULTIPLE COUNTRIES IN AFRICA, EUROPE, ASIA, AND AMERICAS. FOR EXAMPLE, SINCE THE INTRODUCTION OF ASFV IN 2018 IN CHINA, AN ESTIMATED ~50% OF ALL PIGS WERE WIPED OUT. SINCE JULY 2021, ASF HAS BEEN IDENTIFIEDIN THE DOMINICAN REPUBLIC AND VERY RECENTLY IN HAITI, RAPIDLY SPREADING INTO THE CARIBBEAN'S. BECAUSE MOST AMERICAN COUNTRIES ARE NOT WELL PREPARED, INCLUDING MEXICO AND SEVERAL OTHERS COUNTRIES THAT HAVE STRONG COMMERCIAL TRADE OR IMMIGRATION RELATIONSHIPS WITH THE U.S., THE U.S. IS CURRENTLY AT HIGH RISK FOR ASF INTRODUCTION. AS THE WORLD'S THIRD LARGEST PIG PRODUCER, WITH OVER 11.5 MILLION TONS OF PORK PRODUCED PER YEAR, AND THE WORLD'S SECOND LARGEST PORK EXPORTER, THE U.S. MAY FACE FAR-REACHING ECONOMIC CONSEQUENCES IF ASF IS INTRODUCED.TYPICAL ASF DIAGNOSIS REQUIRES IDENTIFICATION OF ANIMALS THAT ARE OR HAVE PREVIOUSLY BEEN INFECTED WITH THE ASF VIRUS (ASFV). GIVEN THE ABSENCE OF TREATMENT AND VACCINES, RAPID AND ACCURATE ASF DIAGNOSTICS IS CRUCIAL TO BORDER INSPECTION, SURVEILLANCE, AND OUTBREAK CONTAINMENT. AN APPROPRIATE DIAGNOSIS INVOLVES THE DETECTION AND IDENTIFICATION OF ASFV-SPECIFIC NUCLEIC ACIDS, ANTIGENS, AND/OR ANTIBODIES. CONVENTIONAL TESTS, SUCH AS REAL TIME POLYMER CHAIN REACTION (PCR) AND ENZYME-LINKED IMMUNOSORBENT ASSAYS (ELISA), ALTHOUGH HIGHLY SENSITIVE, ARE LIMITED TO LAB USE DUE TO REQUIRED INSTRUMENT, PERSONNEL TRAINING, AND LONG TURNAROUND READOUT TIME. IN COMPARISON, THE USE OF RAPID AND PORTABLE DIAGNOSTIC TESTS IN THE FIELD AND NEXT TO A PEN, WHERE THE INFECTED ANIMAL POPULATIONS ARE, WOULD FACILITATE EARLIER AND MORE EFFECTIVE DISEASE CONTROL. IN ADDITION, REDUCED COST AND SIMPLIFIED OPERATION WILL FACILITATE HIGH-FREQUENCY DEPLOYMENT OF SUCH TESTS FOR LARGE-VOLUME AND IN-TIME SURVEILLANCE, WHICH IS IMPORTANT TO DISRUPTING TRANSMISSION CHAIN AND ASF CONTAINMENT. CURRENTLY, THERE IS STILL A LACK OF LOW-COST YET ACCURATE ANTIBODY OR ANTIGEN TESTS THAT CAN BE DEPLOYED TO PEN SIDES FOR SURVEILLANCE PURPOSES.TO BRIDGE THESE TECHNOLOGICAL GAPS, THE WANG LAB AT ARIZONA STATE UNIVERSITY (ASU) AND THE CENTRO DE INVESTIGACI&OACUTE;N EN SANIDAD ANIMAL (INIA-CISA/CSIC) IN SPAIN, WHICH IS ALSO THE EUROPEAN UNION REFERENCE LABORATORY (EURL) AND THE FOOD AND AGRICULTURE ORGANIZATION (FAO) REFERENCE CENTRE (WRC) FOR ASF, WILL COLLABORATIVELY CARRY OUT THE PROPOSED RESEARCH. OUR LONG-TERM GOAL IS TO INNOVATE PORTABLE ELECTRONIC SENSORS THAT CAN BE DEPLOYED WORLDWIDE TO ASF-PREVALENT AREAS FOR RAPID AND SENSITIVE DETECTION OF ASF-SPECIFIC ANTIBODIES AND ANTIGENS AND TO ASSIST ASF CONTAINMENT IN FUTURE OUTBREAKS. IN THIS PROJECT, OUR OBJECTIVE IS TO DESIGN AND VALIDATE A NEW METAL NANOPARTICLE (MNP)-BASED ASSAY PLATFORM, NAMELY NANOPARTICLE-SUPPORTED RAPID, ELECTRONIC DETECTION (NASRED), AND QUANTIFY ITS PERFORMANCE IN ASF DIAGNOSTICS. THE NASRED ASSAY UTILIZES SETS OF MNPS OF DIFFERENT OPTICAL CHARACTERISTICS TO INTERROGATE THE ASF BIOMARKERS, AND GENERATES OPTICALLY DISTINGUISHABLE SIGNALS THAT SERVE TO DETECT SUCH MARKERS IN A MULTIPLEXED FASHION. HERE, WE WILL SIMULTANEOUSLY DETECT ASF ANTIGENS (E.G. P72) AND ANTIBODIES (E.G. ANTI-P54, ANTI-P30, ETC.) AND EVALUATE ITS FEASIBILITY TO IMPROVE THE DIAGNOSTIC ACCURACY. WE WILL COMBINE STRUCTURAL ANALYSIS AND OPTICAL DETECTION WITH INTUITIVE PHYSICAL AND MATHEMATICAL MODELS TO COMPREHENSIVELY UNDERSTAND THE MECHANISMS OF MNP-BASED MULTIVALENT ANALYTE-BINDING AND SUBSEQUENT CLUSTERING AND PRECIPITATION AT NANOMETER SCALE. FURTHER, WE WILL DEVISE A PORTABLE (COMPARABLE TO A U.S. QUARTER) AND INEXPENSIVE SYSTEM (A FEW DOLLARS IN MATERIAL COST) THAT CAN CONVERT THE OPTICAL SIGNALS FOR ELECTRONIC READOUT USING SIMPLE ELECTRONIC CIRCUITS. ADDITIONALLY, WE WILL PROVE THE FEASIBILITY OF RAPID ASF DETECTION (EXPECTED A FEW MINUTES) AND ASSESS THE ASSAY PERFORMANCE FOR PEN-SIDE FIELD DEPLOYMENT.