Multiplex detection of mutations.
Rapid and reliable detection of mutations at the genetic level is an integral part of modern molecular diagnostics. These mutations can range from dominant single nucleotide polymorphisms within specific loci to codominant heterozygotic insertions and they present considerable challenges to investigators in developing rapid nucleic acid-based amplification assays that can distinguish wild-type from mutant alleles. The recent improvements of real-time polymerase chain reaction (PCR) using self-reporting fluorescence probes have given researchers a powerful tool in developing assays for mutation detection that can be multiplexed for high-throughput screening of multiple mutations and cost effectiveness. Here we describe an application of a multiplexed real-time PCR assay using Molecular Beacon probes for the detection of mutations in codon 54 of the CYP51A gene in Aspergillus fumigatus conferring triazole resistance.
Perlin
David S
DS
Public Health Research Institute, UMDNJ-New Jersey Medical School, International Center for Public Health, Newark, NJ, USA.
Balashov
Sergey
S
Park
Steven
S
D001232:Aspergillus fumigatus / Q000187:drug effects / Q000235:genetics*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D004271:DNA, Fungal / Q000032:analysis; D025141:Drug Resistance, Fungal / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D015345:Oligonucleotide Probes / Q000737:chemistry*; D017354:Point Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D014230:Triazoles / Q000494:pharmacology
D004271:DNA, Fungal; D005656:Fungal Proteins; D015345:Oligonucleotide Probes; D014230:Triazoles; D003577:Cytochrome P-450 Enzyme System; C465526:cytochrome P-450 CYP51A, Aspergillus