BSA reduces inhibition in a TaqMan® assay for the detection of Batrachochytrium dendrobatidis
BSA reduces inhibition in a TaqMan® assay for the detection of Batrachochytrium dendrobatidis
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DOI：
10.3354/dao02053
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发表时间：
2010-11-25
影响因子：
1.4
通讯作者：
Skerratt, Lee F.
中科院分区：
文献类型：
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作者：
Garland, Stephen;Baker, Anthony;Skerratt, Lee F.
两栖动物壶菌病病原体（Batrachochytrium dendrobaerum）的TaqMan（R）测定可被酚类化合物（包括腐殖酸和鞣酸）抑制，导致假阴性。已知当样品被这些抑制剂污染时，牛血清白蛋白（BSA）可降低PCR抑制。当分析皮肤拭子中的B时，我们评估了BSA在降低TaqMan（R）测定的抑制中的作用。树枝状的我们发现向TaqMan反应中加入BSA可将抑制降低到不显著的水平。在分析不含环境抑制剂的标准DNA溶液时，BSA对TaqMan反应的效率或分析灵敏度没有明显影响。我们建议在标准TaqMan（R）测定中加入400 ng mul（-1）BSA，以减少与野生两栖动物取样相关的抑制。
A TaqMan (R) 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 (R) assay when analyzing skin swabs for B. dendrobatidis. We found that the addition of BSA to the TaqMan (R) reaction reduced inhibition to insignificant levels. BSA did not appreciably affect the efficiency or analytical sensitivity of the TaqMan (R) reaction in the analysis of standard DNA solutions free from environmental inhibitors. We recommend the addition of 400 ng mu l(-1) of BSA to the standard TaqMan (R) assay to reduce inhibition associated with sampling wild amphibians.