title: Bat white-nose syndrome: a real-time TaqMan polymerase chain reaction test targeting the intergenic spacer region of Geomyces destructans.
abstract: The fungus Geomyces destructans is the causative agent of white-nose syndrome (WNS), a disease that has killed millions of North American hibernating bats. We describe a real-time TaqMan PCR test that detects DNA from G. destructans by targeting a portion of the multicopy intergenic spacer region of the rRNA gene complex. The test is highly sensitive, consistently detecting as little as 3.3 fg genomic DNA from G. destructans. The real-time PCR test specifically amplified genomic DNA from G. destructans but did not amplify target sequence from 54 closely related fungal isolates (including 43 Geomyces spp. isolates) associated with bats. The test was qualified further by analyzing DNA extracted from 91 bat wing skin samples, and PCR results matched histopathology findings. These data indicate the real-time TaqMan PCR method described herein is a sensitive, specific and rapid test to detect DNA from G. destructans and provides a valuable tool for WNS diagnostics and research.
authors.affiliation: US Geological Survey, National Wildlife Health Center, 6006 Schroeder Road, Madison, Wisconsin 53711, USA.
authors.forename: Laura K
authors.initials: LK
authors.lastname: Muller
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Lorch
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Lindner
authors.forename: Michael
authors.initials: M
authors.lastname: O'Connor
authors.forename: Andrea
authors.initials: A
authors.lastname: Gargas
authors.forename: David S
authors.initials: DS
authors.lastname: Blehert
mesh_terms: D000818:Animals; D001203:Ascomycota / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D002685:Chiroptera / Q000382:microbiology*; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D021901:DNA, Intergenic; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D009181:Mycoses / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D010802:Phylogeny; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity; D012867:Skin / Q000382:microbiology; D013045:Species Specificity; D013997:Time Factors; D014481:United States
chemical_list: D004271:DNA, Fungal; D021901:DNA, Intergenic; D021903:DNA, Ribosomal Spacer