title: Diagnostic assays and sampling protocols for the detection of Batrachochytrium dendrobatidis.
abstract: Batrachochytrium dendrobatidis is a fungus belonging to the Phylum Chytridiomycota, Class Chytridiomycetes, Order Chytridiales, and is the highly infectious aetiological agent responsible for a potentially fatal disease, chytridiomycosis, which is currently decimating many of the world's amphibian populations. The fungus infects 2 amphibian orders (Anura and Caudata), 14 families and at least 200 species and is responsible for at least 1 species extinction. Whilst the origin of the agent and routes of transmission are being debated, it has been recognised that successful management of the disease will require effective sampling regimes and detection assays. We have developed a range of unique sampling protocols together with diagnostic assays for the detection of B. dendrobatidis in both living and deceased tadpoles and adults. Here, we formally present our data and discuss them in respect to assay sensitivity, specificity, repeatability and reproducibility. We suggest that compliance with the recommended protocols will avoid the generation of spurious results, thereby providing the international scientific and regulatory community with a set of validated procedures which will assist in the successful management of chytridiomycosis in the future.
authors.affiliation: Australian Animal Health Laboratory, CSIRO Livestock Industries, Private Bag 24, Geelong, Victoria 3220, Australia. alex.hyatt@csiro.au
authors.forename: A D
authors.initials: AD
authors.lastname: Hyatt
authors.forename: D G
authors.initials: DG
authors.lastname: Boyle
authors.forename: V
authors.initials: V
authors.lastname: Olsen
authors.forename: D B
authors.initials: DB
authors.lastname: Boyle
authors.forename: L
authors.initials: L
authors.lastname: Berger
authors.forename: D
authors.initials: D
authors.lastname: Obendorf
authors.forename: A
authors.initials: A
authors.lastname: Dalton
authors.forename: K
authors.initials: K
authors.lastname: Kriger
authors.forename: M
authors.initials: M
authors.lastname: Heros
authors.forename: H
authors.initials: H
authors.lastname: Hines
authors.forename: R
authors.initials: R
authors.lastname: Phillott
authors.forename: R
authors.initials: R
authors.lastname: Campbell
authors.forename: G
authors.initials: G
authors.lastname: Marantelli
authors.forename: F
authors.initials: F
authors.lastname: Gleason
authors.forename: A
authors.initials: A
authors.lastname: Coiling
mesh_terms: D000818:Animals; D001001:Anura / Q000382:microbiology*; D008411:Chytridiomycota / Q000235:genetics / Q000302:isolation & purification*; D004271:DNA, Fungal / Q000032:analysis; D000431:Ethanol / Q000494:pharmacology; D007124:Immunoenzyme Techniques / Q000662:veterinary; D007814:Larva / Q000382:microbiology; D009181:Mycoses / Q000175:diagnosis / Q000473:pathology / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013172:Spores, Fungal / Q000187:drug effects / Q000302:isolation & purification; D013696:Temperature; D014034:Toes / Q000382:microbiology; D014871:Water Microbiology
chemical_list: D004271:DNA, Fungal; D000431:Ethanol