Item Details

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