{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T14:02:46Z","timestamp":1789394566330,"version":"build-2803163510"},"reference-count":44,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T00:00:00Z","timestamp":1789171200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"European Union\u2014NextGenerationEU\u2014PNRR","award":["H91I23000050007"],"award-info":[{"award-number":["H91I23000050007"]}]},{"award":["H91I23000050007"],"award-info":[{"award-number":["H91I23000050007"]}],"id":[{"id":"https:\/\/ror.org\/019w4f821","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Progresso Agricolo Fasano s.c.p.a."},{"name":"National Recovery and Resilience Plan","award":["H93C22000440007"],"award-info":[{"award-number":["H93C22000440007"]}]},{"name":"Agritech National Research Centre","award":["D.D. 1032 17\/06\/2022"],"award-info":[{"award-number":["D.D. 1032 17\/06\/2022"]}]},{"name":"Agritech National Research Centre","award":["CN00000022"],"award-info":[{"award-number":["CN00000022"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J. Fungi","JoF"],"abstract":"<jats:p>Cercospora beticola, the causal agent of Cercospora leaf spot (CLS), is a major pathogen affecting beet (Beta vulgaris) worldwide. Rapid and accurate detection is critical for effective disease control. This study validated a diagnostic framework combining portable quantitative PCR (qPCR) and laboratory-based digital droplet PCR (ddPCR) assays to detect and quantify C. beticola in leaves, seeds, and soil-associated crop residues. We used two portable qPCR systems (Biomeme Franklin\u00ae Three9 and Hyris bCUBE\u2122) for in-field diagnostics, and ddPCR (Bio-Rad Laboratories) for laboratory analysis. Two rapid DNA extraction protocols, the Sigma-Aldrich REDExtract-N-Amp Plant PCR Kit (brief incubation at 95 \u00b0C) and the BN QuickPick Plant DNA Kit (magnetic-bead-based), were evaluated. The optimized portable qPCR assays achieved detection limits of 1 pg \u03bcL\u22121, PCR efficiencies of 96\u2013102%, and R2 &gt; 0.98. The ddPCR assay enabled accurate and absolute quantification even at low target concentrations and was resilient to common PCR inhibitors. Both methods proved robust in complex environmental matrices, with a detection threshold of 1% in infected residue soil. The framework\u2019s sensitivity, specificity, and portability support timely on-site CLS diagnosis and informed management. This integrated approach advances portable molecular diagnostics and provides a platform for future field-ready disease surveillance and sustainable crop protection.<\/jats:p>","DOI":"10.3390\/jof12090686","type":"journal-article","created":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T12:09:51Z","timestamp":1789387791000},"page":"686","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Portable qPCR and ddPCR Diagnostics for Cercospora Leaf Spot: Integrating Field and Laboratory Detection of Cercospora beticola in Beet"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-8878-8608","authenticated-orcid":false,"given":"Marco","family":"Crudele","sequence":"first","affiliation":[{"name":"Department of Soil, Plant and Food Sciences, University of Bari Aldo Moro, 70126 Bari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-4082-9214","authenticated-orcid":false,"given":"Mekides Dugo","family":"Bati","sequence":"additional","affiliation":[{"name":"Department of Soil, Plant and Food Sciences, University of Bari Aldo Moro, 70126 Bari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-0149-2074","authenticated-orcid":false,"given":"Sebastiano","family":"Laera","sequence":"additional","affiliation":[{"name":"Department of Soil, Plant and Food Sciences, University of Bari Aldo Moro, 70126 Bari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4953-468X","authenticated-orcid":false,"given":"Cataldo","family":"Laguardia","sequence":"additional","affiliation":[{"name":"Department of Soil, Plant and Food Sciences, University of Bari Aldo Moro, 70126 Bari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6267-5172","authenticated-orcid":false,"given":"Tiziana","family":"Mascia","sequence":"additional","affiliation":[{"name":"Department of Soil, Plant and Food Sciences, University of Bari Aldo Moro, 70126 Bari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1063-0607","authenticated-orcid":false,"given":"Francesco","family":"Faretra","sequence":"additional","affiliation":[{"name":"Department of Soil, Plant and Food Sciences, University of Bari Aldo Moro, 70126 Bari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5130-0076","authenticated-orcid":false,"given":"Rita Milvia","family":"De Miccolis Angelini","sequence":"additional","affiliation":[{"name":"Department of Soil, Plant and Food Sciences, University of Bari Aldo Moro, 70126 Bari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Vogel, J., Kenter, C., Holst, C., and M\u00e4rl\u00e4nder, B. 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