| RRC ID |
87417
|
| 著者 |
Nozue S, Furuhashi K, Toguchi A, Ishikawa J, Nagura O, Yamashita K, Maekawa M, Iwaizumi M.
|
| タイトル |
Rapid antifungal susceptibility testing for Aspergillus fumigatus using a loop-mediated isothermal amplification method.
|
| ジャーナル |
J Microbiol Methods
|
| Abstract |
In patients with hematological diseases and immunodeficiency, aspergillosis is associated with high mortality, and the early selection of effective antifungal agents is critical. Considering the emergence of azole-resistant Aspergillus and the skill- and time-intensive nature of Clinical and Laboratory Standards Institute (CLSI)-compliant broth microdilution (BMD), there is an urgent need for a rapid and simple antifungal susceptibility testing method. Here, we adapted a loop-mediated isothermal amplification (LAMP)-based phenotypic approach to detect azole resistance in Aspergillus fumigatus, using itraconazole (ITCZ) as the index drug. We evaluated 26 isolates (20 ITCZ-susceptible and 6 ITCZ-resistant strains), whose reference minimum inhibitory concentrations (MICs) were determined using CLSI M38 BMD method. The fungi were incubated for 24 h in Roswell Park Memorial Institute medium (RPMI) medium with and without ITCZ (4 μg/mL), followed by LAMP amplification with real-time monitoring. The time to positivity was recorded as the threshold time (Tt), and susceptibility is summarized as ΔTt = Tt (+ITCZ) - Tt (-ITCZ). The Tt values showed a strong negative correlation with fungal suspension concentration (Spearman r = -0.9686, p < 0.0001). Among the clinical and reference isolates, the ΔTt values ranged from +200 to +592 s for susceptible strains and - 14 to +140 s for resistant strains. A receiver operation characteristic (ROC) curve-derived ΔTt cutoff of 150 s perfectly discriminated all isolates (sensitivity 100% [95% confidence interval (CI), 54.1-100.0]; specificity 100% [95% CI, 83.2-100.0]; AUC 1.00). Our approach produced categorical calls after 24 h of incubation and approximately 1 h of assay time, substantially faster than standard BMD. A rapid and simple LAMP-based antifungal susceptibility approach using ΔTt after brief drug exposure accurately identified ITCZ-resistant A. fumigatus.
|
| 巻・号 |
244
|
| ページ |
107455
|
| 公開日 |
2026-5-1
|
| DOI |
10.1016/j.mimet.2026.107455
|
| PII |
S0167-7012(26)00067-9
|
| PMID |
41831694
|
| MeSH |
Antifungal Agents* / pharmacology
Aspergillosis / microbiology
Aspergillus fumigatus* / drug effects
Aspergillus fumigatus* / genetics
Aspergillus fumigatus* / isolation & purification
Drug Resistance, Fungal
Humans
Itraconazole / pharmacology
Microbial Sensitivity Tests / methods
Molecular Diagnostic Techniques* / methods
Nucleic Acid Amplification Techniques* / methods
|
| IF |
1.707
|
| リソース情報 |
| 病原真核微生物 |
Asperrgillus fumigatus strains IFM 60237, IFM 61567, IFM 64159, IFM 64732, IFM 65548 |
