Molecular Characterization and Antifungal Susceptibility Testing of Sequentially Obtained Clinical Cryptococcus deneoformans and Cryptococcus neoformans Isolates from Ljubljana, Slovenia

TY - JOUR

T1 - Molecular Characterization and Antifungal Susceptibility Testing of Sequentially Obtained Clinical Cryptococcus deneoformans and Cryptococcus neoformans Isolates from Ljubljana, Slovenia

AU - Tomazin, Rok

AU - Matos, Tadeja

AU - Meis, Jacques F

AU - Hagen, Ferry

PY - 2018/4

Y1 - 2018/4

N2 - AIM: To retrospectively investigate the epidemiology of cryptococcosis in Ljubljana, Slovenia.METHODOLOGY: Forty-six sequentially obtained isolates from 19 patients were subjected to amplified fragment length polymorphism (AFLP) genotyping, microsatellite typing, mating- and serotype PCRs and antifungal susceptibility testing.RESULTS: Majority of the isolates were Cryptococcus deneoformans (n = 29/46; 63%) followed by Cryptococcus neoformans (n = 16/46; 34.8%) and their interspecies hybrid (n = 1/46; 2.2%). Mating-type α was predominant, two mating-type a C. deneoformans isolates and one mating-type a/α isolate were observed. Several mixed infections were found by microsatellite typing; one patient had a persisting C. deneoformans infection for > 2.5 years. For C. deneoformans, the in vitro antifungal MIC90 and susceptibility ranges were for amphotericin B 0.25 µg/ml (0.031-0.25 µg/ml), 5-fluorocytosine 0.25 µg/ml (0.063-4 µg/ml), fluconazole 8 µg/ml (0.5-16 µg/ml), voriconazole 0.063 µg/ml (0.008-0.125 µg/ml), posaconazole 0.063 µg/ml (0.008-0.063 µg/ml) and itraconazole 0.063 µg/ml (0.031-0.125 µg/ml). For C. neoformans, these values were for amphotericin B 0.25 µg/ml (0.063-0.5 µg/ml), 5-fluorocytosine 1 µg/ml (0.063-1 µg/ml), fluconazole 16 µg/ml (0.5-64 µg/ml), voriconazole 0.125 µg/ml (0.008-0.25 µg/ml), posaconazole 0.063 µg/ml (0.008-0.063 µg/ml) and itraconazole 0.063 µg/ml (0.031-0.125 µg/ml).CONCLUSIONS: Majority of the cases were caused by C. deneoformans; mating-type α was predominant. Several mixed infections were identified by AFLP genotyping and microsatellite typing. Despite antifungal therapy, a cryptococcal isolate could persist for years. Voriconazole, itraconazole and posaconazole were the most potent antifungal drugs.

AB - AIM: To retrospectively investigate the epidemiology of cryptococcosis in Ljubljana, Slovenia.METHODOLOGY: Forty-six sequentially obtained isolates from 19 patients were subjected to amplified fragment length polymorphism (AFLP) genotyping, microsatellite typing, mating- and serotype PCRs and antifungal susceptibility testing.RESULTS: Majority of the isolates were Cryptococcus deneoformans (n = 29/46; 63%) followed by Cryptococcus neoformans (n = 16/46; 34.8%) and their interspecies hybrid (n = 1/46; 2.2%). Mating-type α was predominant, two mating-type a C. deneoformans isolates and one mating-type a/α isolate were observed. Several mixed infections were found by microsatellite typing; one patient had a persisting C. deneoformans infection for > 2.5 years. For C. deneoformans, the in vitro antifungal MIC90 and susceptibility ranges were for amphotericin B 0.25 µg/ml (0.031-0.25 µg/ml), 5-fluorocytosine 0.25 µg/ml (0.063-4 µg/ml), fluconazole 8 µg/ml (0.5-16 µg/ml), voriconazole 0.063 µg/ml (0.008-0.125 µg/ml), posaconazole 0.063 µg/ml (0.008-0.063 µg/ml) and itraconazole 0.063 µg/ml (0.031-0.125 µg/ml). For C. neoformans, these values were for amphotericin B 0.25 µg/ml (0.063-0.5 µg/ml), 5-fluorocytosine 1 µg/ml (0.063-1 µg/ml), fluconazole 16 µg/ml (0.5-64 µg/ml), voriconazole 0.125 µg/ml (0.008-0.25 µg/ml), posaconazole 0.063 µg/ml (0.008-0.063 µg/ml) and itraconazole 0.063 µg/ml (0.031-0.125 µg/ml).CONCLUSIONS: Majority of the cases were caused by C. deneoformans; mating-type α was predominant. Several mixed infections were identified by AFLP genotyping and microsatellite typing. Despite antifungal therapy, a cryptococcal isolate could persist for years. Voriconazole, itraconazole and posaconazole were the most potent antifungal drugs.

KW - Adolescent

KW - Adult

KW - Aged

KW - Aged, 80 and over

KW - Coinfection/epidemiology

KW - Cryptococcosis/epidemiology

KW - Cryptococcus/classification

KW - Drug Resistance, Fungal

KW - Female

KW - Genes, Mating Type, Fungal

KW - Genotype

KW - Humans

KW - Male

KW - Microbial Sensitivity Tests

KW - Microsatellite Repeats

KW - Middle Aged

KW - Molecular Epidemiology

KW - Molecular Typing

KW - Mycological Typing Techniques

KW - Polymerase Chain Reaction

KW - Retrospective Studies

KW - Serogroup

KW - Slovenia/epidemiology

KW - Young Adult

U2 - 10.1007/s11046-017-0214-9

DO - 10.1007/s11046-017-0214-9

M3 - Article

C2 - 29064061

SN - 0301-486X

VL - 183

SP - 371

EP - 380

JO - Mycopathologia

JF - Mycopathologia

IS - 2

ER -