Techniques for minimizing the effects of PCR inhibitors in the chytridiomycosis assay
TA Kosch, K Summers
Molecular Ecology Resources | WILEY-BLACKWELL | Published : 2013
Chytridiomycosis is an amphibian disease of global conservation concern that is caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd). Since the discovery of Bd in 1998, several methods have been used for detection of Bd; among these polymerase chain reaction (PCR) from skin swabs is accepted as the best method due to its noninvasiveness, high sensitivity and ease of use. However, PCR is not without problems - to be successful, this technique is dependent upon the presence of nondegraded DNA template and reaction contents that are free from inhibitors. Here, we report on an investigation of several techniques aimed at improving the reliability of the Bd PCR assay by minimizing th..View full abstract
This investigation was funded by an East Carolina University Professorship Grant awarded to Kyle Summers. We are grateful to John Wood for his advice on this investigation. Joyce Longcore for providing the Batrachochytrium dendrobatidis isolates for our positive controls. We thank Matt Schrenk, Tim Christensen and Trip Lamb for their advice on techniques; and Mary Norris and Kenneth Sisk for laboratory assistance.