DAO 92:113-116 (2010) - DOI: https://doi.org/10.3354/dao02053
Contribution to the DAO Special 'Chytridiomycosis: An emerging disease'
BSA reduces inhibition in a TaqMan® assay for the detection of Batrachochytrium dendrobatidis
Stephen Garland1,*, Anthony Baker2, Andrea D. Phillott1, Lee F. Skerratt1
ABSTRACT: 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.
KEY WORDS: Batrachochytrium dendrobatidis · Chytridiomycosis · TaqMan assay · Quantitative PCR · Inhibition · Bovine serum albumin · BSA
| Full text in pdf format | Cite this article as: Garland S, Baker A, Phillott AD, Skerratt LF
(2010) BSA reduces inhibition in a TaqMan® assay for the detection of Batrachochytrium dendrobatidis. Dis Aquat Org 92:113-116. https://doi.org/10.3354/dao02053
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