BSA reduces inhibition in a TaqMan® assay for the detection of Batrachochytrium dendrobatidis
Garland, Stephen, Baker, Anthony, Phillott, Andrea D., and Skerratt, Lee F. (2010) BSA reduces inhibition in a TaqMan® assay for the detection of Batrachochytrium dendrobatidis. Diseases of Aquatic Organisms, 92 (2-3). pp. 113-116.
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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 ID: | 5392 |
|---|---|
| Item Type: | Article (Research - C1) |
| ISSN: | 1616-1580 |
| Keywords: | Batrachochytrium dendrobatidis; chytridiomycosis; TaqMan assay; quantitative; PCR; inhibition; bovine serum albumin; BSA |
| Date Deposited: | 13 Oct 2009 05:39 |
| FoR Codes: | 07 AGRICULTURAL AND VETERINARY SCIENCES > 0707 Veterinary Sciences > 070703 Veterinary Diagnosis and Diagnostics @ 100% |
| SEO Codes: | 96 ENVIRONMENT > 9608 Flora, Fauna and Biodiversity > 960807 Fresh, Ground and Surface Water Flora, Fauna and Biodiversity @ 100% |
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