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A TaqMan^® assay for the causative agent of chytridiomycosis in amphibians (Batrachochytrium dendrobatidis) can be inhibited by phenolic compounds, including humic and tannic acids, resulting in false negatives. Bovine serum albumin (BSA) is known to reduce inhibition of PCR when samples are contaminated with these inhibitors. We assessed the effect of BSA in reducing inhibition of the TaqMan^® assay when analyzing skin swabs for B. dendrobatidis. We found that the addition of BSA to the TaqMan^® reaction reduced inhibition to insignificant levels. BSA did not appreciably affect the efficiency or analytical sensitivity of the TaqMan^® reaction in the analysis of standard DNA solutions free from environmental inhibitors. We recommend the addition of 400 ng µl^–1 of BSA to the standard TaqMan^® assay to reduce inhibition associated with sampling wild amphibians.

Item Type:	Refereed Article
Keywords:	cytrid, frogs, fungus, Batrachochytrium dendrobatidis
Research Division:	Biological Sciences
Research Group:	Microbiology
Research Field:	Infectious Agents
Objective Division:	Environment
Objective Group:	Environmental and Natural Resource Evaluation
Objective Field:	Environmental and Natural Resource Evaluation not elsewhere classified
UTAS Author:	Baker, A (Dr Anthony Baker)
ID Code:	94952
Year Published:	2010
Web of Science® Times Cited:	49
Deposited By:	Tasmanian Institute of Agriculture
Deposited On:	2014-09-22
Last Modified:	2017-05-04
Downloads:	78 View Download Statistics