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Here, we present an integrated platform that combines a composite bioaerosol collection system with a three-stage enrichment mechanism and recombinase polymerase amplification\u2013CRISPR\/Cas12a (RPA\u2013CRISPR\/Cas12a) detection. The platform incorporates model-based constant-temperature control, stacked optical filters, and multichannel fluorescence imaging. The temperature-control model accurately described the thermal response of the reaction module, while the optical and imaging subsystems supported stable fluorescence readout. Using aerosolized monkeypox virus (MPXV) pseudovirus as a controlled test target, the platform detected target nucleic acid in collection liquid down to 10 copies\/\u03bcL under the tested conditions. These results support the feasibility of integrating aerosol enrichment with isothermal molecular detection in a compact prototype. The main contributions lie in innovations and the engineering integration of bioaerosol collection and enrichment, thermal control of biochemical reactions in molecular detection, and precision fluorescence detection techniques for molecular detection. Regarding molecular detection methods, the biochemical reaction parameters were experimentally optimized rather than introducing new biochemical reaction mechanisms. Validation with authentic clinical or environmental viral aerosols remains necessary before diagnostic or field-use claims can be made.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c01983","type":"journal-article","created":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T20:12:26Z","timestamp":1787343146000},"page":"24876-24887","source":"Crossref","is-referenced-by-count":1,"title":["High-Sensitivity\nMolecular Detection of Viral Bioaerosols\nUsing a Composite Collection System and Optimized RPA\u2013CRISPR\/Cas12a"],"prefix":"10.1021","volume":"98","author":[{"given":"Hai","family":"Fu","sequence":"first","affiliation":[{"name":"Shenzhen Children\u2019s Hospital , , ,","place":["Shenzhen, Guangdong Province, China, 518038"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhigang","family":"Wang","sequence":"additional","affiliation":[{"name":"Shenzhen Children\u2019s Hospital , , ,","place":["Shenzhen, Guangdong Province, China, 518038"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianxin","family":"Guo","sequence":"additional","affiliation":[{"name":"Shenzhen Children\u2019s Hospital , , ,","place":["Shenzhen, Guangdong Province, China, 518038"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tengfei","family":"Wang","sequence":"additional","affiliation":[{"name":"Shenzhen Children\u2019s Hospital , , ,","place":["Shenzhen, Guangdong Province, China, 518038"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyi","family":"Li","sequence":"additional","affiliation":[{"name":"Shenzhen Children\u2019s Hospital Affiliated to Shantou University Medical College , , ,","place":["ShenZhen, China, 518000"]},{"name":"Shantou University Medical College , ,","place":["Shantou, China, 515041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yimin","family":"Su","sequence":"additional","affiliation":[{"name":"Shenzhen Children\u2019s Hospital Affiliated to Shantou University Medical College , , ,","place":["ShenZhen, China, 518000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7624-3143","authenticated-orcid":true,"given":"Zhi","family":"Chen","sequence":"additional","affiliation":[{"name":"Xinjiang Technical Institute of Physics and Chemistry, Chinese Academy of Sciences , ,","place":["Urumqi, China, 830011"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-5857-671X","authenticated-orcid":true,"given":"Huifeng","family":"Yang","sequence":"additional","affiliation":[{"name":"Shenzhen Children\u2019s Hospital , , ,","place":["Shenzhen, Guangdong Province, China, 518038"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhongjian","family":"Xie","sequence":"additional","affiliation":[{"name":"Shenzhen Children\u2019s Hospital , , ,","place":["Shenzhen, Guangdong Province, China, 518038"]},{"name":"Shenzhen Children\u2019s Hospital Affiliated to Shantou University Medical College , , ,","place":["ShenZhen, China, 518000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Defa","family":"Li","sequence":"additional","affiliation":[{"name":"Shenzhen Children\u2019s Hospital , , ,","place":["Shenzhen, Guangdong Province, China, 518038"]},{"name":"Clinical Laboratory of Shenzhen Children\u2019s Hospital & Shenzhen University Medical School , ,","place":["ShenZhen, China, 518055"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,20]]},"reference":[{"issue":"24","key":"2026090306471688200_cit1","doi-asserted-by":"publisher","first-page":"8531","DOI":"10.1039\/D3CS00417A","article-title":"On-site airborne pathogen\ndetection for infection risk mitigation","volume":"52","author":"Qiu","year":"2023","journal-title":"Chem.\nSoc. 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