DAO 97:249-253 (2012)  -  DOI: https://doi.org/10.3354/dao02420

NOTE
Fast quantitative PCR, locked nucleic acid probes and reduced volume reactions are effective tools for detecting Batrachochytrium dendrobatidis DNA

Gregory R. Ruthig1,2,*, Benjamin P. DeRidder1

1Department of Biology, Grinnell College, 1116 8th Avenue, Grinnell, Iowa 50112, USA
2Present address: Department of Biology, North Central College, 30 North Brainard Street, Naperville, Illinois 60540, USA

ABSTRACT: The fungal pathogen Batrachochytrium dendrobatidis threatens amphibian populations around the world. The ability to detect this pathogen on infected animals and in the environment is critical for understanding and controlling this pandemic. We tested several advances in quantitative PCR (qPCR) techniques to detect B. dendrobatidis DNA. We used a fast PCR thermocycler and enzymes that reduced the volume and the duration of the reaction. We also compared a conventional TaqMan minor groove binding (MGB) probe to an identical locked nucleic acid (LNA) counterpart. The fast qPCR reaction had a high degree of sensitivity to B. dendrobatidis DNA. The LNA probe was effective for detecting B. dendrobatidis DNA and produced results ­similar to those of the MGB probe. The modifications that we tested can improve the cost, time efficiency and specificity of quantitative PCR as a tool for detecting pathogen DNA.


KEY WORDS: Batrachochytrium dendrobatidis · Quantitative PCR · Locked nucleic acids · Minor groove binding probe · Amphibian disease


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Cite this article as: Ruthig GR, DeRidder BP (2012) Fast quantitative PCR, locked nucleic acid probes and reduced volume reactions are effective tools for detecting Batrachochytrium dendrobatidis DNA. Dis Aquat Org 97:249-253. https://doi.org/10.3354/dao02420

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