Rapid detection of triazole antifungal resistance in Aspergillus fumigatus.
Triazole resistance in Aspergillus fumigatus is an uncommon but rising phenomenon. Susceptibility testing is rarely performed and can take 48 h or longer, which is an impediment to effective therapy. Molecular diagnostic probing of well-defined resistance mechanisms, which serve as surrogate markers, provides an alternative approach to rapidly (within hours) and efficiently identify resistant strains. The mechanisms of triazole resistance in A. fumigatus are limited to amino acid substitutions in the drug target Cyp51A and include amino acid substitutions at the positions Gly 54, Gly 138, Met 220, and Leu 98, coupled with a tandem repetition in the gene promoter. We report the development of a real-time PCR assay utilizing molecular beacons to assess triazole resistance markers in A. fumigatus. When combined in a multiplex platform, the assay provides a comprehensive evaluation of drug resistance in A. fumigatus.
Garcia-Effron
Guillermo
G
Public Health Research Institute, International Center for Public Health, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, NJ 07103-3535, USA.
Dilger
Amanda
A
Alcazar-Fuoli
Laura
L
Park
Steven
S
Mellado
Emilia
E
Perlin
David S
DS
D019943:Amino Acid Substitution; D000935:Antifungal Agents / Q000494:pharmacology*; D001232:Aspergillus fumigatus / Q000187:drug effects*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D025141:Drug Resistance, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D013997:Time Factors; D014230:Triazoles / Q000494:pharmacology*
D000935:Antifungal Agents; D015342:DNA Probes; D005656:Fungal Proteins; D014230:Triazoles; D003577:Cytochrome P-450 Enzyme System; C465526:cytochrome P-450 CYP51A, Aspergillus