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First application of loop‐mediated isothermal amplification (LAMP) assays for rapid identification of mating type in the heterothallic fungus Aspergillus fumigatus

BACKGROUND: Loop‐mediated isothermal amplification (LAMP) assays, which operate at a single temperature and require no postreaction processing, have been described for rapid species‐specific detection of numerous fungi. The technology has much less commonly been applied to identification of other ke...

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Autores principales: King, Kevin M., Hawkins, Nichola J., Atkins, Sarah, Dyer, Paul S., West, Jonathan S., Fraaije, Bart A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6771684/
https://www.ncbi.nlm.nih.gov/pubmed/31211900
http://dx.doi.org/10.1111/myc.12959
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author King, Kevin M.
Hawkins, Nichola J.
Atkins, Sarah
Dyer, Paul S.
West, Jonathan S.
Fraaije, Bart A.
author_facet King, Kevin M.
Hawkins, Nichola J.
Atkins, Sarah
Dyer, Paul S.
West, Jonathan S.
Fraaije, Bart A.
author_sort King, Kevin M.
collection PubMed
description BACKGROUND: Loop‐mediated isothermal amplification (LAMP) assays, which operate at a single temperature and require no postreaction processing, have been described for rapid species‐specific detection of numerous fungi. The technology has much less commonly been applied to identification of other key genetic traits such as fungicide resistance, and has not yet been applied to mating‐type determination in any fungus. OBJECTIVES: To develop first LAMP assays for mating‐type identification in a fungus, in this instance with the saprophytic mould and human opportunistic pathogen Aspergillus fumigatus, a heterothallic ascomycete requiring isolates of opposite mating type (MAT1‐1, MAT1‐2) for sexual reproduction. METHODS: New LAMP primer sets, targeted to MAT gene sequences, were screened against 34 A fumigatus isolates (of known mating type) from diverse clinical, environmental and geographic sources to establish whether they could distinguish MAT1‐1 or MAT1‐2 genotypes. RESULTS AND CONCLUSIONS: The new assays, operating at a single temperature of 65°C, correctly identified the mating type of A fumigatus isolates in <20 minutes, and thus have numerous research and practical applications. Similar MAT LAMP assays could now be developed for other fungi of agricultural, environmental, industrial and/or medical importance.
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spelling pubmed-67716842019-10-07 First application of loop‐mediated isothermal amplification (LAMP) assays for rapid identification of mating type in the heterothallic fungus Aspergillus fumigatus King, Kevin M. Hawkins, Nichola J. Atkins, Sarah Dyer, Paul S. West, Jonathan S. Fraaije, Bart A. Mycoses Original Articles BACKGROUND: Loop‐mediated isothermal amplification (LAMP) assays, which operate at a single temperature and require no postreaction processing, have been described for rapid species‐specific detection of numerous fungi. The technology has much less commonly been applied to identification of other key genetic traits such as fungicide resistance, and has not yet been applied to mating‐type determination in any fungus. OBJECTIVES: To develop first LAMP assays for mating‐type identification in a fungus, in this instance with the saprophytic mould and human opportunistic pathogen Aspergillus fumigatus, a heterothallic ascomycete requiring isolates of opposite mating type (MAT1‐1, MAT1‐2) for sexual reproduction. METHODS: New LAMP primer sets, targeted to MAT gene sequences, were screened against 34 A fumigatus isolates (of known mating type) from diverse clinical, environmental and geographic sources to establish whether they could distinguish MAT1‐1 or MAT1‐2 genotypes. RESULTS AND CONCLUSIONS: The new assays, operating at a single temperature of 65°C, correctly identified the mating type of A fumigatus isolates in <20 minutes, and thus have numerous research and practical applications. Similar MAT LAMP assays could now be developed for other fungi of agricultural, environmental, industrial and/or medical importance. John Wiley and Sons Inc. 2019-07-10 2019-09 /pmc/articles/PMC6771684/ /pubmed/31211900 http://dx.doi.org/10.1111/myc.12959 Text en © 2019 The Authors. Mycoses Published by Blackwell Verlag GmbH. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
King, Kevin M.
Hawkins, Nichola J.
Atkins, Sarah
Dyer, Paul S.
West, Jonathan S.
Fraaije, Bart A.
First application of loop‐mediated isothermal amplification (LAMP) assays for rapid identification of mating type in the heterothallic fungus Aspergillus fumigatus
title First application of loop‐mediated isothermal amplification (LAMP) assays for rapid identification of mating type in the heterothallic fungus Aspergillus fumigatus
title_full First application of loop‐mediated isothermal amplification (LAMP) assays for rapid identification of mating type in the heterothallic fungus Aspergillus fumigatus
title_fullStr First application of loop‐mediated isothermal amplification (LAMP) assays for rapid identification of mating type in the heterothallic fungus Aspergillus fumigatus
title_full_unstemmed First application of loop‐mediated isothermal amplification (LAMP) assays for rapid identification of mating type in the heterothallic fungus Aspergillus fumigatus
title_short First application of loop‐mediated isothermal amplification (LAMP) assays for rapid identification of mating type in the heterothallic fungus Aspergillus fumigatus
title_sort first application of loop‐mediated isothermal amplification (lamp) assays for rapid identification of mating type in the heterothallic fungus aspergillus fumigatus
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6771684/
https://www.ncbi.nlm.nih.gov/pubmed/31211900
http://dx.doi.org/10.1111/myc.12959
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