Application of deoxyribonucleic acid microarray analysis to the study of azole antifungal resistance in Candida albicans.
The near completion of sequencing the Candida albicans genome has made it possible to employ genomic technologies, such as microarray analysis, to aid in identifying key genes involved in such clinical problems as the acquisition of high-level resistance to azole antifungal agents. Here, we outline in detail the methodologies utilized in our laboratory to culture clinical isolates of C. albicans, isolate ribonucleic acid from such cultures, synthesize labeled complimentary deoxyribonucleic acid probes from the ribonucleic acid samples, hybridize the probes to microarray chips, analyze the data from such hybridizations, and validate results using reverse transcriptase-polymerase chain reactions. Microarray analysis gives researchers the ability to identify genes involved in processes such as acquisition of azole resistance and to use the data in a way that may lead to clinical approaches to inactivate these genes and improve patient outcomes.
Barker
Katherine S
KS
Department of Pharmacy, College of Pharmacy, University of Tennessee Health Science Center, Memphis, USA.
Rogers
P David
PD
D000935:Antifungal Agents / Q000494:pharmacology*; D001393:Azoles / Q000494:pharmacology*; D002176:Candida albicans / Q000187:drug effects* / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D025141:Drug Resistance, Fungal / Q000235:genetics*; D005800:Genes, Fungal; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012331:RNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D000935:Antifungal Agents; D001393:Azoles; D018076:DNA, Complementary; D004271:DNA, Fungal; D012331:RNA, Fungal