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To address this, the phase separation\u2010assisted pre\u2010enrichment (PSAP) technology is presented that exploits differential polymer\u2010polymer partitioning to achieve 47\u2010fold antigen and 44\u2010fold RNA enrichment simultaneously. Through systematic optimization of interfacial chemistry, including pH modulation, polymer hydrophilicity, mass fraction, and molecular weights, the protocol is refined to enable direct integration with commercial diagnostics. PSAP\u2010boosted rapid antigen ests (RATs) detected SARS\u2010CoV\u20102 and Influenza viruses at tenfold and fivefold lower limits, respectively. In clinical validation, 53 clinical specimens (containing PCR undetectable samples as controls) are analyzed. The PSAP method significantly enhanced detection accuracy for both viral antigens and RNA, particularly improving positivity rates in low viral load specimens (27 &lt; Ct &lt; 31) compared to conventional approaches while maintaining specificity in high\u2010Ct and negative controls. With its universality and tunability, PSAP demonstrates universal applicability across respiratory pathogens and lays the foundation for next\u2010generation point\u2010of\u2010care diagnostics.<\/jats:p>","DOI":"10.1002\/advs.202506578","type":"journal-article","created":{"date-parts":[[2025,6,27]],"date-time":"2025-06-27T09:25:19Z","timestamp":1751016319000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["A Phase Separation\u2010Assisted Pre\u2010Enrichment Method for Ultrasensitive Respiratory Virus Detection"],"prefix":"10.1002","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0813-2740","authenticated-orcid":false,"given":"Yang","family":"Cao","sequence":"first","affiliation":[{"name":"Department of Mechanical Engineering The University of Hong Kong  Pokfulam Road Hong Kong SAR 000000 China"},{"name":"Advanced Biomedical Instrumentation Centre  Hong Kong Science Park, Shatin, New Territories Hong Kong SAR 000000 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pui Ngan","family":"Lau","sequence":"additional","affiliation":[{"name":"Division of Public Health Laboratory Sciences School of Public Health Li Ka Shing Faculty of Medicine The University of Hong Kong  Hong Kong SAR 000000 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alex W.H.","family":"Chin","sequence":"additional","affiliation":[{"name":"Division of Public Health Laboratory Sciences School of Public Health Li Ka Shing Faculty of Medicine The University of Hong Kong  Hong Kong SAR 000000 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhuolin","family":"He","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering The University of Hong Kong  Pokfulam Road Hong Kong SAR 000000 China"},{"name":"Weiyang College Tsinghua University  Beijing 100084 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christina C. 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