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All reports consistently identify\n                    <jats:italic>Trichoderma afroharzianum<\/jats:italic>\n                    as the causal agent. The potential impact on maize production, combined with the risk of misdiagnosis as other ear rot diseases, highlights the need for a rapid and accurate diagnostic method for the reliable detection of\n                    <jats:italic>T. afroharzianum<\/jats:italic>\n                    . In this study, we developed a PCR-based diagnostic tool including an endpoint PCR and a real-time TaqMan assay targeting the TEF1\u03b1 and RPB2 loci, respectively. Both assays were validated using a broad panel of\n                    <jats:italic>Trichoderma<\/jats:italic>\n                    isolates and other fungal species obtained from maize plants, additional host species from diverse geographic origins, and soils collected from German maize fields. All performance criteria required to demonstrate assay reliability were met. The two PCR assays demonstrated higher analytical specificity and inclusivity, albeit were unable to distinguish between pathogenic and non-pathogenic\n                    <jats:italic>T. afroharzianum<\/jats:italic>\n                    isolates. They showed high accuracy and sensitivity, with limits of detection of 1 pg \u00b5L\u207b\u00b9 for the endpoint PCR and 1\u00a0fg \u00b5L\u207b\u00b9 for the real-time PCR. The real-time assay exhibited amplification efficiencies of 105,5% (R\u00b2 = 0.977) for pure cultures and 116% (R\u00b2 = 0.997) for infected maize kernels. Furthermore, both assays reliably detected\n                    <jats:italic>T. afroharzianum<\/jats:italic>\n                    in asymptomatic maize cobs following artificial inoculation with pathogenic strains. These methods therefore enable early and reliable detection of\n                    <jats:italic>T. afroharzianum<\/jats:italic>\n                    ear rot and allow its discrimination from other maize ear rot diseases. This capability is essential for the development of targeted management strategies to address this emerging threat to maize production.\n                  <\/jats:p>","DOI":"10.1038\/s41598-026-51199-2","type":"journal-article","created":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T17:54:10Z","timestamp":1778090050000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and validation of conventional and TaqMan real-time PCR for the detection of Trichoderma afroharzianum causing corn ear rot"],"prefix":"10.1038","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7182-5905","authenticated-orcid":false,"given":"Clovis","family":"Douanla-Meli","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annette","family":"Pfordt","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andreas","family":"von Tiedemann","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gritta","family":"Schrader","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bernhard Carl","family":"Sch\u00e4fer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,5,6]]},"reference":[{"key":"51199_CR1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1080\/23311932.2016.1166995","volume":"2","author":"T Rouf Shah","year":"2016","unstructured":"Rouf Shah, T., Prasad, K., Kumar, P. & Maize A potential source of human nutrition and health: A review. 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