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Quantitative, Competitive PCR and Fluorescent Microscopy Methods for the Study of Batrachochytrium dendrobatidis

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title
Quantitative, Competitive PCR and Fluorescent Microscopy Methods for the Study of Batrachochytrium dendrobatidis
author
Friesen, Leon Ray
abstract
Chytridiomycosis is a disease affecting amphibian populations worldwide and is caused by the parasitic fungus Batrachochytrium dendrobatidis (Bd). The catastrophic decline in numbers of amphibians dictates the need for full characterization of the fungus and amphibian host responses. Significant areas of research that would benefit from strengthening available methods are diagnostic techniques and fundamental molecular biology. A quantitative, competitive PCR (QC-PCR) technique is described that will improve cost efficiency of collecting data on the quantification of Bd in infected animals and the environment. QC-PCR adapts conventional PCR reagents and a competitor DNA sequence into a quantitative technique. Fluorescent microscopy is a vital tool used to study cellular and molecular biology. DNA and cell wall fluorescent staining of viable Bd observed with laser scanning confocal microscopy is described and should provide the foundation for future studies of Bd involving fluorescent microscopy. Both QC-PCR and fluorescent microscopy are technique advancements that will contribute to future studies and to understanding of Bd and chytridiomycosis.
subject
Batrachochytrium dendrobatidis
chytridiomycosis
Fluorescent microscopy
QC-PCR
contributor
Kuhn, Raymond E (committee chair)
Esch, Gerald W (committee member)
Johnson, Erik C (committee member)
date
2011-07-14T20:35:30Z (accessioned)
2012-01-14T09:30:12Z (available)
2011 (issued)
degree
Biology (discipline)
embargo
2012-01-14 (terms)
identifier
http://hdl.handle.net/10339/33447 (uri)
language
en (iso)
publisher
Wake Forest University
type
Thesis

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