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

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.

Grantor dc:publisher

Wake Forest University

Year dc:date.issued

2011

Author dc:creator

• Friesen, Leon Ray

• Batrachochytrium dendrobatidis

Language dc:language.iso

en

Handle dc:identifier.uri

http://hdl.handle.net/10339/33447

OAI identifier oai:identifier

oai:wakespace.lib.wfu.edu:10339/33447