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94952 - BSA reduces inhibition in a TaqMan assay.pdf (87.58 kB)

BSA reduces inhibition in a TaqMan® assay for the detection of Batrachochytrium dendrobatidis

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posted on 2023-05-18, 03:33 authored by Garland, 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

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