Rapid Detection of FKS -Associated Echinocandin Resistance in Candida glabrata

A novel and highly accurate diagnostic assay platform was established for rapid identification ofFKS mutations associated with echinocandin resistance inCandida glabrata . The assay platform uses allele-specific molecular beacon and DNA melt analysis following asymmetric PCR. A dual assay forFKS1 andFKS2 was developed to identify within 3 h the most common and clinically relevant resistance-associated mutations, including 8FKS1 HS1 (wild type [WT], S629P, F625S, D632Y, D632E [T1896G], D632E [T1896A], I634V, and F625F) and 7FKS2 HS1 (WT, F659del, F659S, F659V, F659L, S663P, and S663F) genotypes. A blinded panel of 188C. glabrata clinical isolates was tested by both assays. The molecular diagnostic results from the dual assay were 100% concordant with data obtained from DNA sequencing. This platform has the potential to overcome the deficiencies of existingin vitro susceptibility-based assays to identify echinocandin-resistantC. glabrata and holds promise as a surrogate diagnostic method to better direct echinocandin therapy.