Tracking the amphibian pathogens Batrachochytrium dendrobatidis and Batrachochytrium salamandrivorans using a highly specific monoclonal antibody and lateral‐flow technology

Summary The fungus Batrachochytrium dendrobatidis (Bd) causes chytridiomycosis, a lethal epizootic disease of amphibians. Rapid identification of the pathogen and biosecurity is essential to prevent its spread, but current laboratory‐based tests are time‐consuming and require specialist equipment. H...

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Bibliographic Details
Published in:Microbial biotechnology 2017-03, Vol.10 (2), p.381-394
Main Authors: Dillon, Michael J., Bowkett, Andrew E., Bungard, Michael J., Beckman, Katie M., O'Brien, Michelle F., Bates, Kieran, Fisher, Matthew C., Stevens, Jamie R., Thornton, Christopher R.
Format: Article
Language:English
Subjects:
Amphibians
Animals
Antibodies, Fungal - immunology
Antibodies, Monoclonal - immunology
Antigens
Antigens, Fungal - analysis
Anura
Batrachochytrium dendrobatidis
Biodiversity
Biosecurity
Caudata
Chromatography, Affinity - methods
Chytridiomycosis
Chytridiomycota - isolation & purification
Cotton
Deoxyribonucleic acid
Diagnostic Tests, Routine - methods
DNA
Endangered & extinct species
Frogs
Fungi
Glycoproteins
Immunoglobulin M
Immunoglobulin M - immunology
Infections
Laboratories
Monoclonal antibodies
Mycoses - diagnosis
Mycoses - microbiology
Mycoses - veterinary
Oomycetes
Pathogens
Polyrhiza
Reptiles & amphibians
Salamandridae
Skin
Sporangia
Tracking
Urodela - microbiology
Variance analysis
Veterinary Medicine - methods
Zoosporangia
Zoospores
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Summary:Summary The fungus Batrachochytrium dendrobatidis (Bd) causes chytridiomycosis, a lethal epizootic disease of amphibians. Rapid identification of the pathogen and biosecurity is essential to prevent its spread, but current laboratory‐based tests are time‐consuming and require specialist equipment. Here, we describe the generation of an IgM monoclonal antibody (mAb), 5C4, specific to Bd as well as the related salamander and newt pathogen Batrachochytrium salamandrivorans (Bsal). The mAb, which binds to a glycoprotein antigen present on the surface of zoospores, sporangia and zoosporangia, was used to develop a lateral‐flow assay (LFA) for rapid (15 min) detection of the pathogens. The LFA detects known lineages of Bd and also Bsal, as well as the closely related fungus Homolaphlyctis polyrhiza, but does not detect a wide range of related and unrelated fungi and oomycetes likely to be present in amphibian habitats. When combined with a simple swabbing procedure, the LFA was 100% accurate in detecting the water‐soluble 5C4 antigen present in skin, foot and pelvic samples from frogs, newts and salamanders naturally infected with Bd or Bsal. Our results demonstrate the potential of the portable LFA as a rapid qualitative assay for tracking these amphibian pathogens and as an adjunct test to nucleic acid‐based detection methods. The paper describes the development of a monoclonal antibody, 5C4, and lateral‐flow assay (LFA) specific to the amphibian pathogens Batrachochytrium dendrobatidis and Batrachochytrium salamandrivorans. When combined with a simple swabbing procedure, the LFA allows rapid (15 min) detection of diagnostic antigen in frogs, newts and salamanders naturally‐infected with Bd and Bsal, with 100% accuracy of the LFA confirmed by qPCR of tissue samples.
ISSN:1751-7915
1751-7915
DOI:10.1111/1751-7915.12464