| Abstrakt: |
ABSTRACTReal-time quantitative PCR was used to measure expression levels of genes encoding efflux pumps, ERG11and two control genes, ACT1and PMA1, in a collection of 14 fluconazole-susceptible Candida albicansisolates. For each gene, average expression levels and variations within the population were determined. These values were then used as reference points to make predictions about the molecular basis of resistance in 38 clinical isolates (the majority of which were resistant to fluconazole) obtained from 18 patients treated with posaconazole for refractory oropharyngeal candidiasis. For each of the 38 isolates, the expression levels of genes encoding efflux pumps, ERG11and the control genes, were measured as above. Comparison of the two data sets revealed that expression of ACT1and PMA1did not vary significantly between the two sets of isolates. In contrast, MDR1, ERG11, CDR1, and CDR2were overexpressed in 3, 4, 14, and 35, respectively, of the isolates from patients treated with azoles. In addition to these changes, the patient isolates all had at least one and often multiple missense mutations in ERG11. Select ERG11alleles were expressed in Saccharomyces cerevisiae; all of the alleles tested conferred reduced susceptibility to fluconazole. Despite both the increases in pump expression and the ERG11mutations, only one of the patient isolates exhibited a large decrease in posaconazole susceptibility. |
