Rapid detection of azole-resistant Aspergillus fumigatus in clinical and environmental isolates using lab-on-a-chip diagnostic system
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Supporting information
Accepted version
Author(s)
Type
Journal Article
Abstract
Aspergillus fumigatus has widely evolved resistance to the most commonly used class of antifungal chemicals, the azoles. Current methods for identifying azole resistance are time-consuming and depend on specialized laboratories. There is an urgent need for rapid detection of these emerging pathogens at point-of-care to provide the appropriate treatment in the clinic and to improve management of environmental reservoirs to mitigate the spread of antifungal resistance. Our study demonstrates the rapid and portable detection of the two most relevant genetic markers linked to azole resistance, the mutations TR34 and TR46, found in the promoter region of the gene encoding the azole target, cyp51A. We developed a lab-on-a-chip platform consisting of: (1) tandem-repeat loop-mediated isothermal amplification, (2) state-of-the-art complementary metal-oxide-semiconductor microchip technology for nucleic-acid amplification detection and, (3) and a smartphone application for data acquisition, visualization and cloud connectivity. Specific and sensitive detection was validated with isolates from clinical and environmental samples from 6 countries across 5 continents, showing a lower limit-of-detection of 10 genomic copies per reaction in less than 30 minutes. When fully integrated with a sample preparation module, this diagnostic system will enable the detection of this ubiquitous fungus at the point-of-care, and could help to improve clinical decision making, infection control and epidemiological surveillance.
Date Issued
2020-11
Date Acceptance
2020-08-27
Citation
Journal of Clinical Microbiology, 2020, 58 (11), pp.1-11
ISSN
0095-1137
Publisher
American Society for Microbiology
Start Page
1
End Page
11
Journal / Book Title
Journal of Clinical Microbiology
Volume
58
Issue
11
Copyright Statement
© 2020 American Society for
Microbiology. All Rights Reserved https://doi.org/10.1128/ASMCopyrightv2.
Microbiology. All Rights Reserved https://doi.org/10.1128/ASMCopyrightv2.
Sponsor
Imperial College Healthcare NHS Trust- BRC Funding
National Institute for Health Research
National Institute for Health Research
Identifier
https://jcm.asm.org/content/58/11/e00843-20
Grant Number
RDA02
HPRU-2012-10047
HPRU-2012-10047
Subjects
Aspergillus fumigatus
CMOS
LAMP
TR34
TR46
antimicrobial resistance
azole resistance
isothermal amplification
lab-on-a-chip
point-of-care
Microbiology
06 Biological Sciences
07 Agricultural and Veterinary Sciences
11 Medical and Health Sciences
Publication Status
Published
Date Publish Online
2020-09-09