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Detecting Azole-Antifungal Resistance in Aspergillus fumigatus by Pyrosequencing

1
Mycology Reference Centre, Excellence Centre of Medical Mycology (ECMM), Manchester University NHS Foundation Trust-Wythenshawe Hospital, Manchester M23 9LT, UK
2
Division of Infection, Immunity and Respiratory Medicine, School of Biological Sciences, NIHR Manchester Biomedical Research Centre (BRC) at the Manchester Academic Health Science Centre, The University of Manchester, Manchester M23 9LT, UK
3
Department of Infectious Diseases, Manchester University NHS Foundation Trust-Wythenshawe Hospital, Manchester M23 9LT, UK
*
Author to whom correspondence should be addressed.
J. Fungi 2020, 6(1), 12; https://doi.org/10.3390/jof6010012
Received: 13 December 2019 / Revised: 8 January 2020 / Accepted: 8 January 2020 / Published: 10 January 2020
(This article belongs to the Special Issue Molecular Diagnostics of Fungal Infections)
Guidelines on the diagnosis and management of Aspergillus disease recommend a multi-test approach including CT scans, culture, fungal biomarker tests, microscopy and fungal PCR. The first-line treatment of confirmed invasive aspergillosis (IA) consists of drugs in the azole family; however, the emergence of azole-resistant isolates has negatively impacted the management of IA. Failure to detect azole-resistance dramatically increases the mortality rates of azole-treated patients. Despite drug susceptibility tests not being routinely performed currently, we suggest including resistance testing whilst diagnosing Aspergillus disease. Multiple tools, including DNA sequencing, are available to screen for drug-resistant Aspergillus in clinical samples. This is particularly beneficial as a large proportion of IA samples are culture negative, consequently impeding susceptibility testing through conventional methods. Pyrosequencing is a promising in-house DNA sequencing method that can rapidly screen for genetic hotspots associated with antifungal resistance. Pyrosequencing outperforms other susceptibility testing methods due to its fast turnaround time, accurate detection of polymorphisms within critical genes, including simultaneous detection of wild type and mutated sequences, and—most importantly—it is not limited to specific genes nor fungal species. Here we review current diagnostic methods and highlight the potential of pyrosequencing to aid in a diagnosis complete with a resistance profile to improve clinical outcomes. View Full-Text
Keywords: Aspergillus fumigatus; antifungal drug resistance; cyp51A; azole resistance; diagnostics; pyrosequencing Aspergillus fumigatus; antifungal drug resistance; cyp51A; azole resistance; diagnostics; pyrosequencing
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van der Torre, M.H.; Novak-Frazer, L.; Rautemaa-Richardson, R. Detecting Azole-Antifungal Resistance in Aspergillus fumigatus by Pyrosequencing. J. Fungi 2020, 6, 12.

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