Elizabeth A. Bohuski
Jeffrey M. Lorch
David S. Blehert
Michelle Verant
2016
<p><span>The continued spread of white-nose syndrome and its impacts on hibernating bat populations across North America has prompted nationwide surveillance efforts and the need for high-throughput, noninvasive diagnostic tools. Quantitative real-time polymerase chain reaction (qPCR) analysis has been increasingly used for detection of the causative fungus, </span><i>Pseudogymnoascus destructans</i><span>, in both bat- and environment-associated samples and provides a tool for quantification of fungal DNA useful for research and monitoring purposes. However, precise quantification of nucleic acid from</span><i>P. destructans</i><span> is dependent on effective and standardized methods for extracting nucleic acid from various relevant sample types. We describe optimized methodologies for extracting fungal nucleic acids from sediment, guano, and swab-based samples using commercial kits together with a combination of chemical, enzymatic, and mechanical modifications. Additionally, we define modifications to a previously published intergenic spacer–based qPCR test for </span><i>P. destructans</i><span> to refine quantification capabilities of this assay.</span></p>
application/pdf
10.1177/1040638715626963
en
American Association of Veterinary Laboratory Diagnosticians
Optimized methods for total nucleic acid extraction and quantification of the bat white-nose syndrome fungus, <i>Pseudogymnoascus destructans</i>, from swab and environmental samples
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