20.500.12556/DiRROS-19399 Genome-informed design of a LAMP assay for the specific detection of the strain of ‘Candidatus Phytoplasma asteris’ phytoplasma occurring in grapevines in South Africa Grapevine yellows is one of the most damaging phytoplasma-associated diseases worldwide. It is linked to several phytoplasma species, which can vary regionally due to phytoplasma and insect-vector diversity. Specific, rapid, and reliable detection of the grapevine yellows pathogen has an important role in phytoplasma control. The purpose of this study was to develop and validate a specific loop-mediated isothermal amplification (LAMP) assay for detection of a distinct strain of grapevine ‘Candidatus Phytoplasma asteris’ that is present in South Africa, through implementation of a genome-informed test design approach. Several freely available, user-friendly, web-based tools were coupled to design the specific LAMP assays. The criteria for selection of the assays were set for each step of the process, which resulted in four experimentally operative LAMP assays that targeted the ftsH/hflB gene region, specific to the aster yellows phytoplasma strain from South Africa. A real-time PCR was developed, targeting the same genetic region, to provide extensive validation of the LAMP assay. The validated molecular assays are highly specific to the targeted aster yellows phytoplasma strain from South Africa, with good sensitivity and reproducibility. We show a genome-informed molecular test design and an efficient validation approach for molecular tests if reference and sample materials are sparse and hard to obtain. aster yellows fruit LAMP molecular detection pathogendetection phytoplasma Prokaryotes true false true Angleški jezik Ni določen Neznano 2024-07-17 03:57:01 2024-07-17 13:37:17 2024-07-18 03:54:02 0000-00-00 00:00:00 2022 0 0 Ostali avtorji: Marina Dermastia, Johan Burger, Matthew Dickinson, Gerhard Pietersen, Gert Pietersen, and Tanja Dreo; str. 2927-2839 no. 11 Vol. 106 Nov. 2022 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2022-11-01 632 0191-2917 10.1094/PDIS-10-21-2312-RE 131923203 https://doi.org/10.1094/PDIS-10-21-2312-RE 1 1833e477-43cf-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=26763 RAZ_Alic_Spela_2022.pdf RAZ_Alic_Spela_2022.pdf 1 0BE5585CE8CDB158686876841F73CCA5 d2d5bfd396d0bc4367c921afedd805b3ff0d845b7381617b55b4a62691285947 7ed45fb1-4420-11ef-b487-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=26846 Nacionalni inštitut za biologijo 0 0 0