{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:35:53Z","timestamp":1787027753017,"version":"build-2736575974"},"reference-count":22,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T00:00:00Z","timestamp":1769385600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Zhejiang Provincial Natural Science Foundation of China","award":["LQ23H160002"],"award-info":[{"award-number":["LQ23H160002"]}]},{"name":"Innovation Guidance Special Program for New-type Research and Development Institutions in Jiaxing","award":["2025YDZX028"],"award-info":[{"award-number":["2025YDZX028"]}]},{"name":"Collaborative Project with China Resources Research Institute of Science & Technology","award":["ILH24001"],"award-info":[{"award-number":["ILH24001"]}]},{"name":"Yangtze Delta Region Institute of Tsinghua University, Zhejiang","award":["LZZLX23C002"],"award-info":[{"award-number":["LZZLX23C002"]}]},{"name":"Key Research and Development Program of Shihezi City","award":["2024SF08"],"award-info":[{"award-number":["2024SF08"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Candida albicans is a major fungal pathogen associated with vulvovaginal candidiasis, and rapid, sensitive detection remains challenging, particularly in amplification-free formats. Here, we report NaPddCas, a microfluidic-free, droplet-based CRISPR\/Cas12a detection strategy for qualitative identification of Candida albicans DNA. Unlike conventional bulk CRISPR assays, NaPddCas partitions the reaction mixture into vortex-generated polydisperse droplets, enabling spatial confinement of Cas12a activation events and effective suppression of background fluorescence. This compartmentalization substantially enhances detection sensitivity without nucleic acid amplification or microfluidic devices. Using plasmid and genomic DNA templates, NaPddCas achieved reliable detection at concentrations several orders of magnitude lower than bulk CRISPR\/Cas12a reactions. The assay further demonstrated high specificity against non-target bacterial and fungal species and was successfully applied to clinical vaginal secretion samples. Importantly, NaPddCas is designed as a qualitative or semi-qualitative droplet-dependent digital detection method rather than a quantitative digital assay. Owing to its simplicity, sensitivity, and amplification-free workflow, NaPddCas represents a practical approach for laboratory-based screening of Candida albicans infections.<\/jats:p>","DOI":"10.3390\/bios16020072","type":"journal-article","created":{"date-parts":[[2026,1,26]],"date-time":"2026-01-26T16:13:48Z","timestamp":1769444028000},"page":"72","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Rapid and Sensitive Detection of Candida albicans Using Microfluidic-Free Droplet Digital Non-Amplification Dependent CRISPR\/Cas12a Assay"],"prefix":"10.3390","volume":"16","author":[{"given":"Jie","family":"Peng","sequence":"first","affiliation":[{"name":"Women\u2019s Hospital, School of Medicine, Zhejiang University, Hangzhou 310006, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chao","family":"Guo","sequence":"additional","affiliation":[{"name":"Zhejiang Key Laboratory of Multiomics and Molecular Enzymology, Yangtze Delta Region Institute of Tsinghua University, Zhejiang, Jiaxing 314006, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-8003-4628","authenticated-orcid":false,"given":"Ze-Yun","family":"Huang","sequence":"additional","affiliation":[{"name":"Zhejiang Key Laboratory of Multiomics and Molecular Enzymology, Yangtze Delta Region Institute of Tsinghua University, Zhejiang, Jiaxing 314006, China"},{"name":"College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4257-0100","authenticated-orcid":false,"given":"Wen-Fei","family":"Xu","sequence":"additional","affiliation":[{"name":"Zhejiang Key Laboratory of Multiomics and Molecular Enzymology, Yangtze Delta Region Institute of Tsinghua University, Zhejiang, Jiaxing 314006, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xu-Hui","family":"Li","sequence":"additional","affiliation":[{"name":"Zhejiang Key Laboratory of Multiomics and Molecular Enzymology, Yangtze Delta Region Institute of Tsinghua University, Zhejiang, Jiaxing 314006, China"},{"name":"College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e15992","DOI":"10.5812\/jjm.8(4)2015.15992","article-title":"Isolation of different species of Candida in patients with vulvovaginal candidiasis from sari, iran","volume":"8","author":"Hedayati","year":"2015","journal-title":"Jundishapur J. 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