title: Rapid Detection of FKS-Associated Echinocandin Resistance in Candida glabrata.
abstract: A novel and highly accurate diagnostic assay platform was established for rapid identification of FKS mutations associated with echinocandin resistance in Candida glabrata The assay platform uses allele-specific molecular beacon and DNA melt analysis following asymmetric PCR. A dual assay for FKS1 and FKS2 was developed to identify within 3 h the most common and clinically relevant resistance-associated mutations, including 8 FKS1 HS1 (wild type [WT], S629P, F625S, D632Y, D632E [T1896G], D632E [T1896A], I634V, and F625F) and 7 FKS2 HS1 (WT, F659del, F659S, F659V, F659L, S663P, and S663F) genotypes. A blinded panel of 188 C. 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 existing in vitro susceptibility-based assays to identify echinocandin-resistant C. glabrata and holds promise as a surrogate diagnostic method to better direct echinocandin therapy.
authors.affiliation: Public Health Research Institute, New Jersey Medical School-Rutgers Biomedical and Health Sciences, Newark, New Jersey, USA zhaoy1@njms.rutgers.edu perlinds@njms.rutgers.edu.
authors.forename: Yanan
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Public Health Research Institute, New Jersey Medical School-Rutgers Biomedical and Health Sciences, Newark, New Jersey, USA.
authors.forename: Yoji
authors.initials: Y
authors.lastname: Nagasaki
authors.affiliation: Public Health Research Institute, New Jersey Medical School-Rutgers Biomedical and Health Sciences, Newark, New Jersey, USA.
authors.forename: Milena
authors.initials: M
authors.lastname: Kordalewska
authors.affiliation: Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: Ellen G
authors.initials: EG
authors.lastname: Press
authors.affiliation: Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: Ryan K
authors.initials: RK
authors.lastname: Shields
authors.affiliation: Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: M Hong
authors.initials: MH
authors.lastname: Nguyen
authors.affiliation: Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.|VA Healthcare System, Pittsburgh, Pennsylvania, USA.
authors.forename: Cornelius J
authors.initials: CJ
authors.lastname: Clancy
authors.affiliation: Public Health Research Institute, New Jersey Medical School-Rutgers Biomedical and Health Sciences, Newark, New Jersey, USA zhaoy1@njms.rutgers.edu perlinds@njms.rutgers.edu.
authors.forename: David S
authors.initials: DS
authors.lastname: Perlin
mesh_terms: D000483:Alleles; D000935:Antifungal Agents / Q000494:pharmacology; D041221:Candida glabrata / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D002177:Candidiasis / Q000382:microbiology; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics*; D025141:Drug Resistance, Fungal / Q000235:genetics*; D054714:Echinocandins / Q000494:pharmacology*; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005964:Glucosyltransferases / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D015335:Molecular Probes / Q000737:chemistry; D009154:Mutation; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
chemical_list: D000935:Antifungal Agents; D004271:DNA, Fungal; D054714:Echinocandins; D005656:Fungal Proteins; D007527:Isoenzymes; D015335:Molecular Probes; D005964:Glucosyltransferases