Deciphering Aspergillus fumigatus cyp51A-mediated triazole resistance by pyrosequencing of respiratory specimens
| Autor: | Darin Hassan, Rikesh Masania, Malcolm Richardson, Caroline B. Moore, Lilyann Novak-Frazer, David W. Denning, Samuel P Anees-Hill, Riina Rautemaa-Richardson |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Microbiology (medical)
Azoles Antifungal Agents Aspergillus fumigatus/genetics Cytochrome P-450 Enzyme System/genetics Microbial Sensitivity Tests Aspergillosis Aspergillus fumigatus Microbiology Fungal Proteins symbols.namesake Cytochrome P-450 Enzyme System Polymorphism (computer science) Drug Resistance Fungal Genotype medicine AcademicSubjects/MED00740 Humans Pharmacology (medical) Original Research Pharmacology Sanger sequencing Aspergillus Triazoles/pharmacology biology High-Throughput Nucleotide Sequencing Triazoles biology.organism_classification medicine.disease Fungal Proteins/genetics AcademicSubjects/MED00290 Infectious Diseases Real-time polymerase chain reaction symbols Antifungal Agents/pharmacology Pyrosequencing AcademicSubjects/MED00230 |
| Zdroj: | Novak-Frazer, L, Anees-Hill, S P, Hassan, D, Masania, R, Moore, C B, Richardson, M D, Denning, D W & Rautemaa-Richardson, R 2020, ' Deciphering Aspergillus fumigatus cyp51A-mediated triazole resistance by pyrosequencing of respiratory specimens ', Journal of Antimicrobial Chemotherapy, vol. 75, no. 12, pp. 3501-3509 . https://doi.org/10.1093/jac/dkaa357 Journal of Antimicrobial Chemotherapy |
| DOI: | 10.1093/jac/dkaa357 |
| Popis: | Background Infections caused by triazole drug-resistant Aspergillus fumigatus are an increasing problem. The sensitivity of standard culture is poor, abrogating susceptibility testing. Early detection of resistance can improve patient outcomes, yet tools for this purpose are limited. Objectives To develop and validate a pyrosequencing technique to detect resistance-conferring cyp51A polymorphisms from clinical respiratory specimens and A. fumigatus isolates. Methods Method validation was performed by Sanger sequencing and pyrosequencing of 50 A. fumigatus isolates with a spectrum of triazole susceptibility patterns. Then, 326 Aspergillus quantitative PCR (qPCR)-positive respiratory samples collected over a 27 month period (January 2017–March 2019) from 160 patients at the UK National Aspergillosis Centre were assessed by cyp51A pyrosequencing. The Sanger sequencing and pyrosequencing results were compared with those from high-volume culture and standard susceptibility testing. Results The cyp51A genotypes of the 50 isolates analysed by pyrosequencing and Sanger sequencing matched. Of the 326 Aspergillus qPCR-positive respiratory specimens, 71.2% were reported with no A. fumigatus growth. Of these, 56.9% (132/232) demonstrated a WT cyp51A genotype and 31.5% (73/232) a resistant genotype by pyrosequencing. Pyrosequencing identified the environmental TR34/L98H mutation in 18.7% (61/326) of the samples in contrast to 6.4% (21/326) pan-azole resistance detected by culture. Importantly, pyrosequencing detected resistance earlier than culture in 23.3% of specimens. Conclusions The pyrosequencing assay described could detect a wide range of cyp51A polymorphisms associated with triazole resistance, including those not identified by commercial assays. This method allowed prompt recognition of resistance and the selection of appropriate antifungal treatment when culture was negative. |
| Databáze: | OpenAIRE |
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