Molecular Identification and Susceptibility Testing of Molds Isolated in a Prospective Surveillance of Triazole Resistance in Spain (FILPOP2 Study)

Abstract

Antifungal resistance is increasing by the emergence of intrinsically resistant species and by the development of secondary resistance in susceptible species. A previous study performed in Spain revealed levels of azole resistance in molds of between 10 and 12.7%, but secondary resistance in <i>Aspergillus fumigatus</i> was not detected. We used itraconazole (ITZ)-supplemented medium to select resistant strains. A total of 500 plates supplemented with 2 mg/liter of ITZ were sent to 10 Spanish tertiary hospitals, and molecular identification and antifungal susceptibility testing were performed. In addition, the <i>cyp51A</i> gene in those <i>A. fumigatus</i> strains showing azole resistance was sequenced. A total of 493 isolates were included in the study. Sixteen strains were isolated from patients with an infection classified as proven, 104 were isolated from patients with an infection classified as probable, and 373 were isolated from patients with an infection classified as colonization. <i>Aspergillus</i> was the most frequent genus isolated, at 80.3%, followed by <i>Scedosporium-Lomentospora</i> (7.9%), <i>Penicillium-Talaromyces</i> (4.5%), <i>Fusarium</i> (2.6%), and the order <i>Mucorales</i> (1%). Antifungal resistance was detected in <i>Scedosporium-Lomentospora</i> species, <i>Fusarium</i>, <i>Talaromyces</i>, and <i>Mucorales</i> Three strains of <i>A. fumigatus sensu stricto</i> were resistant to azoles; two of them harbored the TR₃₄+L98H mechanism of resistance, and the other one had no mutations in <i>cyp51A</i> The level of azole resistance in <i>A. fumigatus</i> remains low, but cryptic species represent over 10% of the isolates and have a broader but overall higher range of antifungal resistance.