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Protocol for the Detection of Chytrid Fungus Using Quantitative PCR Techinques

In terms of its effect on biodiversity, chytridiomycosis is quite possibly the worst disease in recorded history. The disease, caused by a pathogenic chytrid fungus, has caused amphibian population declines in Australia, South America, North America, Central America, New Zealand, Europe, and Africa, and is likely responsible for over 100 species extinctions since the 1970’s.

While chytridiomycosis has become a major focus of amphibian research worldwide, few herpetologists have any background in the laboratory techniques necessary to detect the chytrid fungus, and training classes are rare. Quantitative PCR (qPCR) is the most technologically advanced method of diagnosing chytrid infections, and the ability to perform qPCR in one’s own laboratory would both expedite the processing of samples and decrease the costs incurred by sending samples to private laboratories for diagnosis.

SAVE THE FROGS! has now made publicly and freely available a detailed protocol for the detection and quantification of Batrachochytrium dendrobatidis. This is the most in-depth protocol of its kind in existence, and can be found, along with accompanying slideshow, videos and supplementary materials at:
http://savethefrogs.com/chytrid/qpcr.html

chytrid pcr

The slideshow, videos, webpage and other materials were developed as part of a free course offered by SAVE THE FROGS! and the Herpetological Circle of Panama. The course, entitled “Instruction and application of quantitative PCR molecular techniques for the study of amphibian epidemics”, took place at the Smithsonian Tropical Research Institute in Panama City on October 5th-9th 2009, and was attended by 25 scientists from Panama, Colombia, and Costa Rica. Amphibian population in all three countries have experienced severe declines in numbers due to the chytrid fungus. This course was taught in Spanish by SAVE THE FROGS! Founder & Executive Director Dr. Kerry Kriger, with the valuable assistance of Vicky Flechas of Colombia’s Universidad de Los Andes. This course effectively doubled the number of scientists on the planet capable of using quantitative PCR for the diagnosis of amphibian chytrid infections.

We plan to hold another such course in Bogota, Colombia in November of 2010.
http://savethefrogs.com/chytrid/qpcr.html

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