Rapid Detection of Azole-Resistant Aspergillus fumigatus in Clinical and Environmental Isolates by Use of a Lab-on-a-Chip Diagnostic System
| Autor: | Nicolas Moser, Ling-Shan Yu, Alison Holmes, Johanna Rhodes, Ahmad Moniri, Matthew C. Fisher, Nicholas Miscourides, Amelie P Brackin, Kenny Malpartida-Cardenas, Tatiana Kochina, Thomas R. Sewell, Pantelis Georgiou, Jesus Rodriguez-Manzano |
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| Přispěvatelé: | Imperial College Healthcare NHS Trust- BRC Funding, National Institute for Health Research |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Microbiology (medical) Azoles isothermal amplification Antifungal Agents 030106 microbiology Loop-mediated isothermal amplification Azole resistance Computational biology Mycology Microbial Sensitivity Tests Diagnostic system Rapid detection Microbiology Aspergillus fumigatus law.invention Fungal Proteins 03 medical and health sciences TR34 Antibiotic resistance azole resistance law LAMP Drug Resistance Fungal 07 Agricultural and Veterinary Sciences Lab-On-A-Chip Devices Aspergillosis Humans antimicrobial resistance 11 Medical and Health Sciences chemistry.chemical_classification biology lab-on-a-chip CMOS Lab-on-a-chip 06 Biological Sciences biology.organism_classification TR46 030104 developmental biology chemistry Molecular Diagnostic Techniques point-of-care Mutation Azole Nucleic Acid Amplification Techniques |
| Zdroj: | J Clin Microbiol |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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