Sequential Induction of Drug Resistance and Characterization of an Initial Drug-Sensitive Isolate.

BACKGROUND

The pathogenic fungus is a leading agent of death in immunocompromised individuals with a growing trend of antifungal resistance.

METHODS

The purpose is to induce resistance to drugs in a sensitive strain followed by whole-genome sequencing to determine mechanisms of resistance. Strains will be assayed for pathogenicity attributes such as ergosterol and chitin content, growth rate, virulence, and biofilm formation.

RESULTS

We observed sequential increases in ergosterol and chitin content in fluconazole-resistant isolates by 78% and 44%. Surface thickening prevents the entry of the drug, resulting in resistance. Resistance imposed a fitness trade-off that led to reduced growth rates, biofilm formation, and virulence in our isolates. Sequencing revealed mutations in genes involved in resistance and pathogenicity such as , , , , , and . We observed an increase in the number of mutations in key genes with a sequential increase in drug-selective pressures as the organism increased its odds of adapting to inhospitable environments. In , we observed two mutations in the susceptible strain and five mutations in the resistant strain.

CONCLUSION

This is the first study to induce resistance followed by genotypic and phenotypic analysis of isolates to determine mechanisms of drug resistance.