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BSA reduces inhibition in a TaqMan® assay for the detection of Batrachochytrium dendrobatidis
Citation
Garland, S and Baker, A and Phillott, AD and Skerratt, LF, BSA reduces inhibition in a TaqMan assay for the detection of Batrachochytrium dendrobatidis, Diseases of Aquatic Organisms, 92, (2-3) pp. 113-116. ISSN 0177-5103 (2010) [Refereed Article]
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Copyright Statement
Copyright 2010 Inter-Research
DOI: doi:10.3354/dao02053
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.
Item Details
| Item Type: | Refereed Article |
|---|---|
| Keywords: | cytrid, frogs, fungus, Batrachochytrium dendrobatidis |
| Research Division: | Biological Sciences |
| Research Group: | Microbiology |
| Research Field: | Infectious agents |
| Objective Division: | Environmental Management |
| Objective Group: | Other environmental management |
| Objective Field: | Other environmental management not elsewhere classified |
| UTAS Author: | Baker, A (Dr Anthony Baker) |
| ID Code: | 94952 |
| Year Published: | 2010 |
| Web of Science® Times Cited: | 57 |
| Deposited By: | Tasmanian Institute of Agriculture |
| Deposited On: | 2014-09-22 |
| Last Modified: | 2017-05-04 |
| Downloads: | 110 View Download Statistics |
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