posted on 2023-05-18, 03:33authored byGarland, S, Baker, A, Phillott, AD, Skerratt, LF
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.
History
Publication title
Diseases of Aquatic Organisms
Volume
92
Issue
2-3
Pagination
113-116
ISSN
0177-5103
Department/School
Tasmanian Institute of Agriculture (TIA)
Publisher
Inter-Research
Place of publication
Nordbunte 23, Oldendorf Luhe, Germany, D-21385
Rights statement
Copyright 2010 Inter-Research
Repository Status
Open
Socio-economic Objectives
Other environmental management not elsewhere classified