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

A - Papers appearing in refereed journals

King, K. M., Hawkins, N. J., Atkins, S. L., Dyer, P. S., West, J. S. and Fraaije, B. A. 2019. First application of loop-mediated isothermal amplification (LAMP) assays for rapid identification of mating type in the heterothallic fungus Aspergillus fumigatus . Mycoses.

AuthorsKing, K. M., Hawkins, N. J., Atkins, S. L., Dyer, P. S., West, J. S. and Fraaije, B. A.
Abstract

Background Loop-mediated isothermal amplification (LAMP) assays, which operate at a single temperature and require no post-reaction 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 if 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.

KeywordsAspergillus fumigatus; Diagnostics; Fungal pathogen; Mating type; Sexual reproduction
Year of Publication2019
JournalMycoses
Digital Object Identifier (DOI)doi:10.1111/myc.12959
Web address (URL)https://onlinelibrary.wiley.com/doi/abs/10.1111/myc.12959
Open accessPublished as ‘gold’ (paid) open access
Funder project or codeBBSRC Strategic Programme in Smart Crop Protection
FunderBBSRC Industrial Strategy Challenge
Biotechnology and Biological Sciences Research Council
Accepted author manuscriptsubmission r1..pdf
Output statusPublished
Publication dates
Online18 Jun 2019
Publication process dates
Accepted13 Jun 2019
Copyright licenseCC BY
PublisherWiley
ISSN0933-7407

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