PROJECT SUMMARY
Candida auris is an emerging pan-multidrug-resistant fungal pathogen and presents a serious global health
threat. Importantly, C. auris is the first fungal pathogen showing pronounced and sometimes untreatable clinical
drug resistance to all known antifungal classes, including echinocandins azoles, and amphotericin B.
However, the diagnostic tools for the early detection of drug resistance in C. auris infections are either inadequate
or not sensitive enough for the identification of drug resistant clinical isolates. Thus, the combination of antifungal
drug resistance, along with a lack of sensitive diagnostic assays impair efficient therapeutic strategies to
overcome or prevent drug resistance in C. auris. Hence, the overall objective is to develop and validate
SuperSelective PCR assays for the detection of mutations in genes encoding echinocandin or azole resistance,
despite an abundant background of wild-type C. auris DNA. We hypothesize that SuperSelective PCR assays
will detect mutant C. auris with a sensitivity exceeding 1:104 mutant:wild type for single nucleotide polymorphisms
at genetic loci conferring echinocandin or azole resistance. The premise is based on our proof-of-concept work
with an C. auris FKS1 mutation conferring high-level echinocandin resistance. We expect to test our hypothesis
and to accomplish the objectives by pursuing two Specific Aims: 1) Develop and validate a novel SuperSelective
PCR platform to detect mutations in C. auris genes conferring resistance to echinocandin and azole antifungal
drugs. 2) Validate SuperSelective PCR assays to detect echinocandin and azole heteroresistance in the kidney
homogenates of mice with active candidiasis. The payoffs of this proposal are expected to be significant, because
the proposed study will develop a novel SuperSelective primer PCR platform to identify mutations,
heteroresistant or tolerant clinical C. auris isolates. It will also provide a framework for mechanistic understanding
of multidrug resistance in C. auris.