title: Rapid detection of pathogenic fungi from clinical specimens using LightCycler real-time fluorescence PCR.
abstract: In the study presented here a LightCycler real-time PCR system was used for the diagnosis of fungal infections from clinical tissue samples. Nine specimens were investigated from six patients with suspected or proven invasive fungal infections. Seven of nine samples were positive in a broad-range fungal PCR assay. In four samples, Aspergillus fumigatus was detected both by a species-specific hybridization assay as well as by sequencing of amplification products. In addition, the broad-range fungal PCR assay and PCR sequencing detected and identified, respectively, the following organisms in the specimens noted: Candida albicans in a culture-negative liver biopsy, Histoplasma capsulatum in a bone marrow sample, and Conidiobolus coronatus in a facial soft tissue specimen. Real-time PCR is a promising tool for the diagnosis of invasive fungal infections in human tissue samples and offers some advantages over culture methods, such as rapid analysis and increased sensitivity.
authors.affiliation: Program in Infectious Disease, Clinical Research Division, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue N, D3-100, WA 98109-1024, Seattle, USA.
authors.forename: A
authors.initials: A
authors.lastname: Imhof
authors.forename: C
authors.initials: C
authors.lastname: Schaer
authors.forename: G
authors.initials: G
authors.lastname: Schoedon
authors.forename: D J
authors.initials: DJ
authors.lastname: Schaer
authors.forename: R B
authors.initials: RB
authors.lastname: Walter
authors.forename: A
authors.initials: A
authors.lastname: Schaffner
authors.forename: M
authors.initials: M
authors.lastname: Schneemann
mesh_terms: D001232:Aspergillus fumigatus / Q000302:isolation & purification; D001483:Base Sequence; D002176:Candida albicans / Q000302:isolation & purification; D004271:DNA, Fungal / Q000032:analysis; D005260:Female; D016469:Fungemia / Q000175:diagnosis* / Q000382:microbiology; D005658:Fungi / Q000302:isolation & purification*; D006658:Histoplasma / Q000302:isolation & purification; D006801:Humans; D017403:In Situ Hybridization; D008297:Male; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009181:Mycoses / Q000175:diagnosis* / Q000382:microbiology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012494:Sampling Studies; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D004271:DNA, Fungal