In vitro antifungal susceptibilities and molecular typing of sequentially isolated clinical Cryptococcus neoformans strains from Croatia

Abstract

A collection of 48 clinical Cryptococcus neoformans isolates from Croatia was investigated retrospectively using in vitro antifungal susceptibility testing and molecular biological techniques to determine mating type and serotype by PCR and amplified fragment length polymorphism (AFLP) genotyping. These isolates were obtained from 15 patients: ten were human immunodeficiency virus (HIV)-negative (66.7 %) and five were HIV-positive (33.3 %). From five patients, only one isolate was available, whilst from the other ten patients, two to 11 isolates were isolated sequentially. Antifungal susceptibility was tested by a broth microdilution method. Serotype A (genotype AFLP1) and serotype D (genotype AFLP2) were both found in six patients (40 % each), and serotype AD (genotype AFLP3) in three (20.0 %) patients. Mating type a (n512; 80.0 %) predominated and a/a hybrids were identified in 20.0 % of patients diagnosed with cryptococcosis. Two AFLP genotypes of C. neoformans were isolated during a single episode from one patient. The in vitro antifungal MIC 90 and susceptibility ranges for C. neoformans isolates were 0.5 mg ml "1 (range 0.031-0.5 mg ml ) for flucytosine and fluconazole, 0.25 mg ml "1 (range 0.031-0.5 mg ml "1 ) for itraconazole and 0.062 mg ml "1 (range 0.031-0.25 mg ml "1 ) for voriconazole.

Extensive Genetic Diversity within the Dutch Clinical Cryptococcus neoformans Population

“…The present study shows that genotype AFLP1 and AFLP3 isolates are less susceptible to fluconazole than AFLP2 isolates. This genotypic difference was only recently observed in a Croatian study that found a similar result for fluconazole, although this difference was not significant between AFLP2 and AFLP3 isolates (31). The current study shows that there is a trend toward lower MIC values of amphotericin B for genotype AFLP2 isolates, similarly to that observed in the Croatian and Spanish studies (20,31).…”

“…The current study shows that there is a trend toward lower MIC values of amphotericin B for genotype AFLP2 isolates, similarly to that observed in the Croatian and Spanish studies (20,31). The Croatian and Spanish studies did not observe a significant difference between any of the tested antifungal compounds versus the AFLP genotypes, except for Spanish AFLP1 isolates that were less susceptible to amphotericin B than Spanish AFLP3 isolates (20,31). Similarly to our study, the Spanish study did not observe a significant difference between patient categories versus antifungal susceptibility.…”

“…This genotypic difference was only recently observed in a Croatian study that found a similar result for fluconazole, although this difference was not significant between AFLP2 and AFLP3 isolates (31). The current study shows that there is a trend toward lower MIC values of amphotericin B for genotype AFLP2 isolates, similarly to that observed in the Croatian and Spanish studies (20,31). The Croatian and Spanish studies did not observe a significant difference between any of the tested antifungal compounds versus the AFLP genotypes, except for Spanish AFLP1 isolates that were less susceptible to amphotericin B than Spanish AFLP3 isolates (20,31).…”

Two Rare Cases Of Central Nervous System Opportunistic Mycoses / Oportunističke Mikoze Središnjega Živčanog Sustava – Prikaz Dvaju Bolesnika

“…In Croatia, nearly 900 HIV-infected persons had been registered from 1985, when HIV was first diagnosed, to December 2010 (13). The widespread use of highly active antiretroviral therapy (HAART) has lowered the incidence of cryptococcosis in HIVinfected persons in medically developed countries and in Croatia (11,13).…”

“…The widespread use of highly active antiretroviral therapy (HAART) has lowered the incidence of cryptococcosis in HIVinfected persons in medically developed countries and in Croatia (11,13). Between 2005 and 2010, RCMDSDI verifi ed cryptococcosis in fi ve (33.3 %) HIV-infected patients and ten (66.7 %) non-HIVinfected patients (13).…”