Development of a loop-mediated isothermal amplification assay for the detection of Tilletia controversa based on genome comparison

Abstract Tilletia controversa causing dwarf bunt of wheat is a quarantine pathogen in several countries. Therefore, its specific detection is of great phytosanitary importance. Genomic regions routinely used for phylogenetic inferences lack suitable polymorphisms for the development of species-speci...

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Autores principales:	Somayyeh Sedaghatjoo, Monika K. Forster, Ludwig Niessen, Petr Karlovsky, Berta Killermann, Wolfgang Maier
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4f848145611243da9e6bcd2da7fbeee6
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spelling	oai:doaj.org-article:4f848145611243da9e6bcd2da7fbeee62021-12-02T18:24:54ZDevelopment of a loop-mediated isothermal amplification assay for the detection of Tilletia controversa based on genome comparison10.1038/s41598-021-91098-22045-2322https://doaj.org/article/4f848145611243da9e6bcd2da7fbeee62021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-91098-2https://doaj.org/toc/2045-2322Abstract Tilletia controversa causing dwarf bunt of wheat is a quarantine pathogen in several countries. Therefore, its specific detection is of great phytosanitary importance. Genomic regions routinely used for phylogenetic inferences lack suitable polymorphisms for the development of species-specific markers. We therefore compared 21 genomes of six Tilletia species to identify DNA regions that were unique and conserved in all T. controversa isolates and had no or limited homology to other Tilletia species. A loop-mediated isothermal amplification (LAMP) assay for T. controversa was developed based on one of these DNA regions. The specificity of the assay was verified using 223 fungal samples comprising 43 fungal species including 11 Tilletia species, in particular 39 specimens of T. controversa, 92 of T. caries and 40 of T. laevis, respectively. The assay specifically amplified genomic DNA of T. controversa from pure cultures and teliospores. Only Tilletia trabutii generated false positive signals. The detection limit of the LAMP assay was 5 pg of genomic DNA per reaction. A test performance study that included five laboratories in Germany resulted in 100% sensitivity and 97.7% specificity of the assay. Genomic regions, specific to common bunt (Tilletia caries and Tilletia laevis together) are also provided.Somayyeh SedaghatjooMonika K. ForsterLudwig NiessenPetr KarlovskyBerta KillermannWolfgang MaierNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Somayyeh Sedaghatjoo Monika K. Forster Ludwig Niessen Petr Karlovsky Berta Killermann Wolfgang Maier Development of a loop-mediated isothermal amplification assay for the detection of Tilletia controversa based on genome comparison
description	Abstract Tilletia controversa causing dwarf bunt of wheat is a quarantine pathogen in several countries. Therefore, its specific detection is of great phytosanitary importance. Genomic regions routinely used for phylogenetic inferences lack suitable polymorphisms for the development of species-specific markers. We therefore compared 21 genomes of six Tilletia species to identify DNA regions that were unique and conserved in all T. controversa isolates and had no or limited homology to other Tilletia species. A loop-mediated isothermal amplification (LAMP) assay for T. controversa was developed based on one of these DNA regions. The specificity of the assay was verified using 223 fungal samples comprising 43 fungal species including 11 Tilletia species, in particular 39 specimens of T. controversa, 92 of T. caries and 40 of T. laevis, respectively. The assay specifically amplified genomic DNA of T. controversa from pure cultures and teliospores. Only Tilletia trabutii generated false positive signals. The detection limit of the LAMP assay was 5 pg of genomic DNA per reaction. A test performance study that included five laboratories in Germany resulted in 100% sensitivity and 97.7% specificity of the assay. Genomic regions, specific to common bunt (Tilletia caries and Tilletia laevis together) are also provided.
format	article
author	Somayyeh Sedaghatjoo Monika K. Forster Ludwig Niessen Petr Karlovsky Berta Killermann Wolfgang Maier
author_facet	Somayyeh Sedaghatjoo Monika K. Forster Ludwig Niessen Petr Karlovsky Berta Killermann Wolfgang Maier
author_sort	Somayyeh Sedaghatjoo
title	Development of a loop-mediated isothermal amplification assay for the detection of Tilletia controversa based on genome comparison
title_short	Development of a loop-mediated isothermal amplification assay for the detection of Tilletia controversa based on genome comparison
title_full	Development of a loop-mediated isothermal amplification assay for the detection of Tilletia controversa based on genome comparison
title_fullStr	Development of a loop-mediated isothermal amplification assay for the detection of Tilletia controversa based on genome comparison
title_full_unstemmed	Development of a loop-mediated isothermal amplification assay for the detection of Tilletia controversa based on genome comparison
title_sort	development of a loop-mediated isothermal amplification assay for the detection of tilletia controversa based on genome comparison
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/4f848145611243da9e6bcd2da7fbeee6
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Development of a loop-mediated isothermal amplification assay for the detection of Tilletia controversa based on genome comparison

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