20.500.12556/DiRROS-22173 Test performance study on qPCR assays for detection of Phyllosticta citricarpa Citrus black spot (CBS), caused by the fungus Phyllosticta citricarpa, significantly affects citrus fruit marketability and can lead to premature fruit drop. Accurate and reliable detection of this quarantine pathogen is crucial, particularly for asymptomatic plant material. This study evaluated two qPCR assays, the EPPO recommended assay PC and assay Pc-TEF1, based on TEF region, for detecting P. citricarpa through a collaborative test performance study (TPS). DNA from the isolates of Phyllosticta spp. and other fungi was spiked into citrus fruit peel extracts (lemon, orange, and pomelo) and distributed among 13 laboratories. Sample and qPCR assay stability under typical transport conditions was confirmed, although prolonged storage affected Pc-TEF1 assay performance. The assays were assessed based on sensitivity, specificity, reproducibility, and repeatability. Both assays demonstrated high performance, with repeatability and reproducibility exceeding 95%. The PC assay, as expected, detected different related Phyllosticta species, while Pc-TEF1 showed higher specificity for P. citricarpa included in the TPS alone. Additionally, inhibitory effects were observed specifically in the pomelo peel samples, suggesting matrix-dependent variability. This TPS confirms that both PC and Pc-TEF1 qPCR assays are robust. Further evaluation of the qPCR assays would support the selection of the most reliable assays for the detection of P. citricarpa, contributing to the effective management of CBS disease in citrus production and trade. test performance study Phyllosticta citricarpa real time PCR TEF1 biotechnology true false true Angleški jezik Ni določen Neznano 2025-05-07 09:10:21 2025-05-07 09:10:21 2025-10-24 03:50:42 0000-00-00 00:00:00 2025 0 0 Nasl. z nasl. zaslona; Opis vira z dne 25. 4. 2025; str. 1-12 no. 5, [article no.] 413 Vol. 14 2025 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2025-04-24 581 2076-0817 10.3390/pathogens14050413 234340611 https://doi.org/10.3390/pathogens14050413 1 5952d5bb-2b12-11f0-963a-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=32081 RAZ_Jakomin_Tjasa_2025.pdf RAZ_Jakomin_Tjasa_2025.pdf 1 EEBC582D7DFD7640A66E892A5112C3AE 98a5a6e7850dccda30ad071f2799e92a49f9a241e2cdd2cc697018c0ae4e6807 614185d6-2b12-11f0-963a-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=32082 pathogens-14-00413-s001.zip pathogens-14-00413-s001.zip Dopolnilne informacije Supplementary materials 1 54E95F7DFF9D5D61C3D09D26CB461F03 c072d4327dbd728b1de75d69698fbe260993792ff40f0332988f7616d06ac44b aa2bdce3-2bff-11f0-963a-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=32121 Nacionalni inštitut za biologijo 0 0 0