BSA reduces inhibition in a TaqMan assay for the detection of Batrachochytrium dendrobatidis.
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 microl(-1) of BSA to the standard TaqMan assay to reduce inhibition associated with sampling wild amphibians.
Amphibian Disease Ecology Group, James Cook University, and the School of Public Health, Tropical Medicine and Rehabilitation Sciences, James Cook University, Townsville, Queensland 4811, Australia. stephen.garland@jcu.edu.au
Stephen
S
Garland
Anthony
A
Baker
Andrea D
AD
Phillott
Lee F
LF
Skerratt
D000818:Animals; D008411:Chytridiomycota / Q000235:genetics / Q000302:isolation & purification*; D006812:Humic Substances / Q000032:analysis*; D009181:Mycoses / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary; D010636:Phenols; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011898:Ranidae; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D013634:Tannins / Q000737:chemistry*
D006812:Humic Substances; D010636:Phenols; D013634:Tannins; D012710:Serum Albumin, Bovine