Title |
Optimized methods for total nucleic acid extraction and quantification of the bat white-nose syndrome fungus, Pseudogymnoascus destructans, from swab and environmental samples
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Published in |
Journal of Veterinary Diagnostic Investigation, March 2016
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DOI | 10.1177/1040638715626963 |
Pubmed ID | |
Authors |
Michelle L. Verant, Elizabeth A. Bohuski, Jeffery M. Lorch, David S. Blehert |
Abstract |
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. |
X Demographics
Geographical breakdown
Country | Count | As % |
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Australia | 1 | 100% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 1 | 2% |
Unknown | 45 | 98% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 10 | 22% |
Student > Master | 8 | 17% |
Student > Doctoral Student | 4 | 9% |
Student > Ph. D. Student | 4 | 9% |
Other | 4 | 9% |
Other | 9 | 20% |
Unknown | 7 | 15% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 18 | 39% |
Environmental Science | 7 | 15% |
Veterinary Science and Veterinary Medicine | 3 | 7% |
Biochemistry, Genetics and Molecular Biology | 2 | 4% |
Arts and Humanities | 1 | 2% |
Other | 6 | 13% |
Unknown | 9 | 20% |