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However, most current diagnostics are limited to centralized laboratories or require specialized equipment such as electrochemical readers, with a few hours to multi\u2010day turnaround times that hinder field deployment. Here, we introduce a DNA nanostructure\u2010enabled lateral flow assay (LFA) for rapid, ultrasensitive detection of porcine epidemic diarrhea virus (PEDV) directly on farms. This assay employs net\u2010shaped DNA nanostructures presenting PEDV\u2010specific aptamers in a multivalent configuration, yielding \u223c1000\u2010fold greater binding avidity than free aptamers. The platform demonstrates the limit of detection\u00a0(LOD)\u00a0of \u223c1.33\u00a0ng\/mL for recombinant PEDV nucleocapsid protein and \u223c278 viral copies per test in pig oral fluid samples, with results obtained within 10 min without requiring nucleic acid extraction or amplification. Validation with animal clinical samples (n = 102)\u00a0from swine farms demonstrates an accuracy of 97.06%, sensitivity of 96.92%, and specificity of 97.30% (Ct \u2264 37.42), without cross\u2010reactivity against other swine viruses. This work presents a robust and field\u2010deployable diagnostic platform that combines the programmability of DNA nanotechnology with the operational simplicity of LFA. Beyond PEDV, the modular nature of the DNA\u2010Net platform is readily adaptable for detecting other veterinary and zoonotic pathogens, advancing disease surveillance, and One Health diagnostics.<\/jats:p>","DOI":"10.1002\/adhm.202504037","type":"journal-article","created":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T10:52:38Z","timestamp":1782211958000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Ultrasensitive Detection of Porcine Epidemic Diarrhea Virus Infections Using Multivalent DNA Nanostructure\u2010Enabled Lateral Flow Assay"],"prefix":"10.1002","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9735-8062","authenticated-orcid":false,"given":"Saurabh","family":"Umrao","sequence":"first","affiliation":[{"name":"Department of Bioengineering University of Illinois at Urbana\u2010Champaign  Urbana Illinois USA"},{"name":"Nick Holonyak Jr. Micro and Nanotechnology Laboratory University of Illinois at Urbana\u2010Champaign  Urbana Illinois USA"},{"name":"Carl R. Woese Institute for Genomic Biology University of Illinois at Urbana\u2010Champaign  Urbana Illinois USA"},{"name":"Department of Chemistry University of Illinois at Urbana\u2010Champaign  Urbana Illinois USA"},{"name":"Cancer Center at Illinois University of Illinois at Urbana\u2010Champaign  Urbana Illinois USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akhila","family":"Naru","sequence":"additional","affiliation":[{"name":"Department of Pathobiology College of Veterinary Medicine University of Illinois at Urbana\u2010Champaign  Urbana Illinois USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-3063-4408","authenticated-orcid":false,"given":"Dhanush","family":"Gandavadi","sequence":"additional","affiliation":[{"name":"Department of Bioengineering University of Illinois at Urbana\u2010Champaign  Urbana Illinois USA"},{"name":"Nick Holonyak Jr. Micro and Nanotechnology Laboratory University of Illinois at Urbana\u2010Champaign  Urbana Illinois USA"},{"name":"Carl R. 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