Set of classical PCRs for detection of mutations in Candida glabrata FKS genes linked with echinocandin resistance.
Clinical echinocandin resistance among Candida glabrata strains is increasing, especially in the United States. Antifungal susceptibility testing is considered mandatory to guide therapeutic decisions. However, these methodologies are not routinely performed in the hospital setting due to their complexity and the time needed to obtain reliable results. Echinocandin failure in C. glabrata is linked exclusively to Fks1p and Fks2p amino acid substitutions, and detection of such substitutions would serve as a surrogate marker to identify resistant isolates. In this work, we report an inexpensive, simple, and quick classical PCR set able to objectively detect the most common mechanisms of echinocandin resistance in C. glabrata within 4 h. The usefulness of this assay was assessed using a blind collection of 50 C. glabrata strains, including 16 FKS1 and/or FKS2 mutants.
Laboratorio de Micología y Diagnóstico Molecular, Cátedra de Parasitología y Micología, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe, Argentina Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Santa Fe, Argentina.
Catiana
C
Dudiuk
Laboratorio de Micología y Diagnóstico Molecular, Cátedra de Parasitología y Micología, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe, Argentina.
Soledad
S
Gamarra
Laboratorio de Micología y Diagnóstico Molecular, Cátedra de Parasitología y Micología, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe, Argentina.
Florencia
F
Leonardeli
Public Health Research Institute, Rutgers, The State University of New Jersey, Newark, New Jersey, USA.
Cristina
C
Jimenez-Ortigosa
Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Santa Fe, Argentina Unidad de Parasitología, Laboratorio de Micología, Hospital Ramos Mejía, CONICET, Ciudad Autónoma de Buenos Aires, Argentina.
Roxana G
RG
Vitale
Unidad de Parasitología, Laboratorio de Micología, Hospital Ramos Mejía, CONICET, Ciudad Autónoma de Buenos Aires, Argentina Catedra 1, Departamento de Microbiología, Parasitología e Inmunología, Universidad Nacional de Buenos Aires, Argentina.
Javier
J
Afeltra
Public Health Research Institute, Rutgers, The State University of New Jersey, Newark, New Jersey, USA.
David S
DS
Perlin
Laboratorio de Micología y Diagnóstico Molecular, Cátedra de Parasitología y Micología, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe, Argentina Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Santa Fe, Argentina ggarcia@unl.edu.ar.
Guillermo
G
Garcia-Effron
D019943:Amino Acid Substitution; D000935:Antifungal Agents / Q000494:pharmacology*; D041221:Candida glabrata / Q000187:drug effects* / Q000235:genetics*; D025141:Drug Resistance, Fungal; D054714:Echinocandins / Q000494:pharmacology*; D005656:Fungal Proteins / Q000235:genetics; D005964:Glucosyltransferases / Q000235:genetics*; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods; D025202:Molecular Diagnostic Techniques / Q000379:methods; D050505:Mutant Proteins / Q000235:genetics; D020125:Mutation, Missense; D016133:Polymerase Chain Reaction / Q000379:methods*; D014481:United States
D000935:Antifungal Agents; D054714:Echinocandins; D005656:Fungal Proteins; D050505:Mutant Proteins; D005964:Glucosyltransferases