title: Molecular epidemiology and in vitro antifungal susceptibility testing of 108 clinical Cryptococcus neoformans sensu lato and Cryptococcus gattii sensu lato isolates from Denmark.
abstract: Cryptococcosis is mainly caused by members of the Cryptococcus gattii/Cryptococcus neoformans species complexes. Here, we report the molecular characterisation and in vitro antifungal susceptibility of Danish clinical cryptococcal isolates. Species, genotype, serotype and mating type were determined by amplified fragment length polymorphism (AFLP) fingerprinting and qPCR. EUCAST E.Def 7.2 MICs were determined for amphotericin B, flucytosine, fluconazole, voriconazole and isavuconazole. Most isolates were C. neoformans (serotype A; n = 66) and belonged to genotype AFLP1/VNI (n = 61) or AFLP1B/VNII (n = 5) followed by Cryptococcus deneoformans (serotype D; genotype AFLP2, n = 20), C. neoformans × C. deneoformans hybrids (serotype AD; genotype AFLP3, n = 13) and Cryptococcus curvatus (n = 2). Six isolates were C. gattii sensu lato, and one isolate was a C. deneoformans × C. gattii hybrid (genotype AFLP8). All isolates were amphotericin B susceptible. Flucytosine susceptibility was uniform MIC50 of 4-8 mg l(-1) except for C. curvatus (MICs >32 mg l(-1) ). Cryptococcus gattii sensu lato isolates were somewhat less susceptible to the azoles. MICs of fluconazole (>32 mg l(-1) ), voriconazole (≥0.5 mg l(-1) ) and isavuconazole (0.06 and 0.25 mg l(-1) respectively) were elevated compared to the wild-type population for 1/19 C. deneoformans and 1/2 C. curvatus isolates. Flucytosine MIC was elevated for 1/61 C. neoformans (>32 mg l(-1) ). Antifungal susceptibility revealed species-specific differential susceptibility, but suggested acquired resistance was an infrequent phenomenon.
authors.affiliation: Department of Medical Microbiology and Infectious Diseases, Canisius-Wilhelmina Hospital, Nijmegen, The Netherlands.
authors.forename: Ferry
authors.identifier: http://orcid.org/0000-0002-5622-1916
authors.initials: F
authors.lastname: Hagen
authors.affiliation: Department of Microbiological Surveillance and Research, Unit of Mycology, Statens Serum Institut, Copenhagen, Denmark.
authors.forename: Rasmus
authors.initials: R
authors.lastname: Hare Jensen
authors.affiliation: Department of Medical Microbiology and Infectious Diseases, Canisius-Wilhelmina Hospital, Nijmegen, The Netherlands.|Department of Medical Microbiology, Radboudumc, Nijmegen, The Netherlands.
authors.forename: Jacques F
authors.initials: JF
authors.lastname: Meis
authors.affiliation: Department of Microbiological Surveillance and Research, Unit of Mycology, Statens Serum Institut, Copenhagen, Denmark.
authors.forename: Maiken Cavling
authors.initials: MC
authors.lastname: Arendrup
mesh_terms: D000666:Amphotericin B / Q000494:pharmacology; D000935:Antifungal Agents / Q000494:pharmacology*; D001393:Azoles / Q000494:pharmacology; D003453:Cryptococcosis / Q000453:epidemiology* / Q000382:microbiology; D056285:Cryptococcus gattii / Q000187:drug effects* / Q000235:genetics*; D003455:Cryptococcus neoformans / Q000187:drug effects* / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics; D003718:Denmark / Q000453:epidemiology; D025141:Drug Resistance, Fungal; D015725:Fluconazole / Q000494:pharmacology; D005838:Genotype; D008826:Microbial Sensitivity Tests; D058885:Multilocus Sequence Typing; D010802:Phylogeny; D012703:Serotyping
chemical_list: D000935:Antifungal Agents; D001393:Azoles; D004271:DNA, Fungal; D000666:Amphotericin B; D015725:Fluconazole
keywords: Cryptococcus bacillisporus; Cryptococcus deneoformans; Cryptococcus deuterogattii; Cryptococcus gattii; Cryptococcus neoformans; amplified fragment length polymorphism fingerprinting; antifungal susceptibility testing; multilocus sequence typing