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Mycology

Rapid Detection of Azole-Resistant Aspergillus fumigatus in Clinical and Environmental Isolates by Use of a Lab-on-a-Chip Diagnostic System

Ling-Shan Yu, Jesus Rodriguez-Manzano, Nicolas Moser, Ahmad Moniri, Kenny Malpartida-Cardenas, Nicholas Miscourides, Thomas Sewell, Tatiana Kochina, Amelie Brackin, Johanna Rhodes, Alison H. Holmes, Matthew C. Fisher, Pantelis Georgiou
Kimberly E. Hanson, Editor
Ling-Shan Yu
aCentre for Bio-Inspired Technology, Department of Electrical and Electronic Engineering, Faculty of Engineering, Imperial College London, London, United Kingdom
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Jesus Rodriguez-Manzano
aCentre for Bio-Inspired Technology, Department of Electrical and Electronic Engineering, Faculty of Engineering, Imperial College London, London, United Kingdom
bNIHR Health Protection Research Unit in Healthcare Associated Infections and Antimicrobial Resistance, Department of Infectious Disease, Faculty of Medicine, Imperial College London, London, United Kingdom
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Nicolas Moser
aCentre for Bio-Inspired Technology, Department of Electrical and Electronic Engineering, Faculty of Engineering, Imperial College London, London, United Kingdom
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Ahmad Moniri
aCentre for Bio-Inspired Technology, Department of Electrical and Electronic Engineering, Faculty of Engineering, Imperial College London, London, United Kingdom
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Kenny Malpartida-Cardenas
aCentre for Bio-Inspired Technology, Department of Electrical and Electronic Engineering, Faculty of Engineering, Imperial College London, London, United Kingdom
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Nicholas Miscourides
aCentre for Bio-Inspired Technology, Department of Electrical and Electronic Engineering, Faculty of Engineering, Imperial College London, London, United Kingdom
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Thomas Sewell
cMRC Centre for Global Infectious Disease Analysis, Faculty of Medicine, Imperial College London, London, United Kingdom
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  • ORCID record for Thomas Sewell
Tatiana Kochina
cMRC Centre for Global Infectious Disease Analysis, Faculty of Medicine, Imperial College London, London, United Kingdom
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Amelie Brackin
cMRC Centre for Global Infectious Disease Analysis, Faculty of Medicine, Imperial College London, London, United Kingdom
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Johanna Rhodes
cMRC Centre for Global Infectious Disease Analysis, Faculty of Medicine, Imperial College London, London, United Kingdom
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  • ORCID record for Johanna Rhodes
Alison H. Holmes
bNIHR Health Protection Research Unit in Healthcare Associated Infections and Antimicrobial Resistance, Department of Infectious Disease, Faculty of Medicine, Imperial College London, London, United Kingdom
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Matthew C. Fisher
cMRC Centre for Global Infectious Disease Analysis, Faculty of Medicine, Imperial College London, London, United Kingdom
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Pantelis Georgiou
aCentre for Bio-Inspired Technology, Department of Electrical and Electronic Engineering, Faculty of Engineering, Imperial College London, London, United Kingdom
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Kimberly E. Hanson
University of Utah
Roles: Editor
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DOI: 10.1128/JCM.00843-20
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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: (i) tandem-repeat loop-mediated isothermal amplification; (ii) state-of-the-art complementary metal-oxide-semiconductor microchip technology for nucleic acid amplification detection; and (iii) 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 min. 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.

FOOTNOTES

    • Received 22 April 2020.
    • Returned for modification 16 June 2020.
    • Accepted 27 August 2020.
    • Accepted manuscript posted online 9 September 2020.
  • Supplemental material is available online only.

  • Copyright © 2020 American Society for Microbiology.

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Rapid Detection of Azole-Resistant Aspergillus fumigatus in Clinical and Environmental Isolates by Use of a Lab-on-a-Chip Diagnostic System
Ling-Shan Yu, Jesus Rodriguez-Manzano, Nicolas Moser, Ahmad Moniri, Kenny Malpartida-Cardenas, Nicholas Miscourides, Thomas Sewell, Tatiana Kochina, Amelie Brackin, Johanna Rhodes, Alison H. Holmes, Matthew C. Fisher, Pantelis Georgiou
Journal of Clinical Microbiology Oct 2020, 58 (11) e00843-20; DOI: 10.1128/JCM.00843-20

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Rapid Detection of Azole-Resistant Aspergillus fumigatus in Clinical and Environmental Isolates by Use of a Lab-on-a-Chip Diagnostic System
Ling-Shan Yu, Jesus Rodriguez-Manzano, Nicolas Moser, Ahmad Moniri, Kenny Malpartida-Cardenas, Nicholas Miscourides, Thomas Sewell, Tatiana Kochina, Amelie Brackin, Johanna Rhodes, Alison H. Holmes, Matthew C. Fisher, Pantelis Georgiou
Journal of Clinical Microbiology Oct 2020, 58 (11) e00843-20; DOI: 10.1128/JCM.00843-20
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KEYWORDS

antimicrobial resistance
Aspergillus fumigatus
azole resistance
CMOS
isothermal amplification
LAMP
lab-on-a-chip
point-of-care
TR34
TR46

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