{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T17:50:34Z","timestamp":1774979434400,"version":"3.50.1"},"reference-count":24,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T00:00:00Z","timestamp":1774915200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Natural Science Research Project for Anhui Universities of the Key Scientific Research Foundation of the Education Department of Province Anhui","award":["KJ2021ZD0150"],"award-info":[{"award-number":["KJ2021ZD0150"]}]},{"name":"Research Start-up Fund of Suzhou University of Science and Technology"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>Respiratory fungal infections are challenging to diagnose due to non-specific symptoms and limitations of conventional methods. This study established an integrated, rapid workflow for detecting three common pathogenic fungi, combining an optimized nucleic acid extraction method with a TaqMan-based multiplex real-time qPCR system. The high-salt lysis method yielded high-purity nucleic acid extracts, with performance comparable to or better than commercial kits. The multiplex qPCR assay was optimized for high specificity, sensitivity, and reproducibility, with detection limits as low as 10 CFU\/mL for some fungi. Clinical validation using 14 sputum samples demonstrated high agreement with conventional culture, and the assay detected a low-load infection missed by culture. This integrated approach shows great potential for rapid, sensitive, and specific clinical detection of respiratory fungal infections.<\/jats:p>","DOI":"10.3390\/jof12040252","type":"journal-article","created":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T16:34:08Z","timestamp":1774974848000},"page":"252","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["An Efficient Workflow for Fungal Nucleic Acid Detection in Sputum"],"prefix":"10.3390","volume":"12","author":[{"given":"Ying","family":"Han","sequence":"first","affiliation":[{"name":"School of Chemistry and Life Sciences, Suzhou University of Science and Technology, Suzhou 215009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Deding","family":"Tang","sequence":"additional","affiliation":[{"name":"The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi\u2019an Jiaotong University, Xi\u2019an 710049, China"},{"name":"Bioinspired Engineering and Biomechanics Center (BEBC), Xi\u2019an Jiaotong University, Xi\u2019an 710049, China"},{"name":"Public Teaching Department, Maanshan Teacher\u2019s College, Maanshan 243041, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuntong","family":"Gao","sequence":"additional","affiliation":[{"name":"Suzhou Diyinan Biotech Company, Suzhou 215129, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lilian","family":"Ji","sequence":"additional","affiliation":[{"name":"School of Chemistry and Life Sciences, Suzhou University of Science and Technology, Suzhou 215009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1109-3110","authenticated-orcid":false,"given":"Jie","family":"Hu","sequence":"additional","affiliation":[{"name":"Suzhou Diyinan Biotech Company, Suzhou 215129, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yan","family":"Gong","sequence":"additional","affiliation":[{"name":"School of Chemistry and Life Sciences, Suzhou University of Science and Technology, Suzhou 215009, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1038\/s41579-023-00861-x","article-title":"The WHO fungal priority pathogens list as a game-changer","volume":"21","author":"Fisher","year":"2023","journal-title":"Nat. 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