title: Development and use of a real-time polymerase chain reaction assay for the detection of Ophidiomyces ophiodiicola in snakes.
abstract: Fungal pathogens threatening the conservation of wildlife are becoming increasingly common. Since 2008, free-ranging snakes across North America have been experiencing a marked increase in the prevalence of snake fungal disease associated with Ophidiomyces ophiodiicola. Diagnosis has historically relied on histology, microbiology, and conventional polymerase chain reaction (PCR). More sensitive methods are needed to adequately characterize the epidemiology. The current study describes the development of a real-time PCR (qPCR) assay for detecting a segment of the internal transcribed spacer 1 region between the 18S and 5.8S ribosomal RNA gene. The assay was able to detect as few as 1.05 × 10(1) gene copies per reaction. An additional 4 positive cases were detected when comparing a conventional PCR (n = 3) and the qPCR (n = 7) when used on swab samples from 47 eastern massasauga rattlesnakes. The newly developed assay is a sensitive and specific tool for surveillance and monitoring in the conservation of free-ranging snakes.
authors.affiliation: Departments of Comparative Biosciences (Allender, Bunick, Dzhaman), College of Veterinary Medicine, University of Illinois, Urbana, ILPathobiology (Maddox), College of Veterinary Medicine, University of Illinois, Urbana, ILVeterinary Diagnostic Laboratory (Burrus), College of Veterinary Medicine, University of Illinois, Urbana, IL mattallender@vetmed.illinois.edu.
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Allender
authors.affiliation: Departments of Comparative Biosciences (Allender, Bunick, Dzhaman), College of Veterinary Medicine, University of Illinois, Urbana, ILPathobiology (Maddox), College of Veterinary Medicine, University of Illinois, Urbana, ILVeterinary Diagnostic Laboratory (Burrus), College of Veterinary Medicine, University of Illinois, Urbana, IL.
authors.forename: David
authors.initials: D
authors.lastname: Bunick
authors.affiliation: Departments of Comparative Biosciences (Allender, Bunick, Dzhaman), College of Veterinary Medicine, University of Illinois, Urbana, ILPathobiology (Maddox), College of Veterinary Medicine, University of Illinois, Urbana, ILVeterinary Diagnostic Laboratory (Burrus), College of Veterinary Medicine, University of Illinois, Urbana, IL.
authors.forename: Elena
authors.initials: E
authors.lastname: Dzhaman
authors.affiliation: Departments of Comparative Biosciences (Allender, Bunick, Dzhaman), College of Veterinary Medicine, University of Illinois, Urbana, ILPathobiology (Maddox), College of Veterinary Medicine, University of Illinois, Urbana, ILVeterinary Diagnostic Laboratory (Burrus), College of Veterinary Medicine, University of Illinois, Urbana, IL.
authors.forename: Lucienne
authors.initials: L
authors.lastname: Burrus
authors.affiliation: Departments of Comparative Biosciences (Allender, Bunick, Dzhaman), College of Veterinary Medicine, University of Illinois, Urbana, ILPathobiology (Maddox), College of Veterinary Medicine, University of Illinois, Urbana, ILVeterinary Diagnostic Laboratory (Burrus), College of Veterinary Medicine, University of Illinois, Urbana, IL.
authors.forename: Carol
authors.initials: C
authors.lastname: Maddox
mesh_terms: D000818:Animals; D000835:Animals, Wild; D001203:Ascomycota / Q000302:isolation & purification*; D004271:DNA, Fungal / Q000032:analysis; D003881:Dermatomycoses / Q000382:microbiology / Q000662:veterinary*; D009656:North America; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D012911:Snakes / Q000382:microbiology*
chemical_list: D004271:DNA, Fungal
keywords: Epidemiology; molecular biology; reptile; snake fungal disease