期刊
JOURNAL OF VETERINARY DIAGNOSTIC INVESTIGATION
卷 28, 期 2, 页码 110-118出版社
SAGE PUBLICATIONS INC
DOI: 10.1177/1040638715626963
关键词
Bats; DNA extraction; Pseudogymnoascus destructans; quantification; white-nose syndrome
资金
- U.S. Fish and Wildlife Service [4500036150]
- U.S. Geological Survey
- University of Wisconsin-Madison
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, Pseudogymnoascus destructans, 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 P. destructans 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 P. destructans to refine quantification capabilities of this assay.
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