Use of a high-resolution melt assay to characterize codon 54 of the cyp51A gene of Aspergillus fumigatus on a Rotor-Gene 6000 instrument.
A high-resolution melt (HRM) assay using a Rotor-Gene 6000 instrument was developed to characterize the codon for glycine 54 in the cyp51A genes from 13 reference isolates and 12 clinical isolates of Aspergillus fumigatus. Mutations in this codon confer reduced susceptibility to itraconazole and posaconazole. The assay is simple to perform, and a result of "wild type" or "mutant" is available after approximately 1 h following DNA extraction using commercially available reagents and conventional primers.
Tuohy
M J
MJ
Cleveland Clinic, Cleveland, OH 44195, USA. tuohym@ccf.org
Reja
V
V
Park
S
S
Perlin
D S
DS
Wnek
M
M
Procop
G W
GW
Yen-Lieberman
B
B
D000935:Antifungal Agents / Q000494:pharmacology; D001232:Aspergillus fumigatus / Q000187:drug effects* / Q000235:genetics*; D003062:Codon / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D025141:Drug Resistance, Fungal / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal / Q000235:genetics; D005820:Genetic Testing / Q000379:methods; D005998:Glycine / Q000235:genetics; D017964:Itraconazole / Q000494:pharmacology; D008826:Microbial Sensitivity Tests / Q000379:methods*; D009691:Nucleic Acid Denaturation; D017422:Sequence Analysis, DNA / Q000379:methods; D014230:Triazoles / Q000494:pharmacology
D000935:Antifungal Agents; D003062:Codon; D005656:Fungal Proteins; D014230:Triazoles; D017964:Itraconazole; C101425:posaconazole; D003577:Cytochrome P-450 Enzyme System; C465526:cytochrome P-450 CYP51A, Aspergillus; D005998:Glycine