title: Real-time PCR for detection of the Aspergillus genus.
abstract: Aspergillus is a genus of mold that has strong indoor sources, including several species capable of acting as opportunistic pathogens. Previous studies suggest that Aspergillus could serve as an indicator for abnormal mold growth or moisture, making it an important genus for environmental monitoring. Here, a quantitative polymerase chain reaction (qPCR, or real-time PCR) assay is presented for Aspergillus. The assay shows good specificity for the genus, detecting all Aspergillus species tested, although a few non-Aspergillus species are also amplified. Sensitivity testing demonstrates that DNA representing one conidium can be detected. A validation study compared qPCR results against direct microscopy counts using A. fumigatus conidia aerosolized into a laboratory chamber. The assay was then used to quantify Aspergillus in indoor air samples, demonstrating its utility for environmental monitoring. Analysis of a small number of clinical sputum samples showed complete agreement with culturing results.
authors.affiliation: Stanford University, Civil and Environmental Engineering Dept, Terman Engineering Center B13, Stanford, CA 94305, USA. mdgoebes@stanfordalumni.org
authors.forename: Marian D
authors.initials: MD
authors.lastname: Goebes
authors.forename: Lynn M
authors.initials: LM
authors.lastname: Hildemann
authors.forename: Elmira
authors.initials: E
authors.lastname: Kujundzic
authors.forename: Mark
authors.initials: M
authors.lastname: Hernandez
mesh_terms: D001230:Aspergillus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D004271:DNA, Fungal / Q000235:genetics; D004784:Environmental Monitoring / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013172:Spores, Fungal / Q000235:genetics
chemical_list: D004271:DNA, Fungal