Optimized methods for total nucleic acid extraction and quantification of the bat white-nose syndrome fungus, Pseudogymnoascus destructans, from swab and environmental samples

Journal of Veterinary Diagnostic Investigation
By: , and 

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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 fromP. 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.

Additional publication details

Publication type Article
Publication Subtype Journal Article
Title Optimized methods for total nucleic acid extraction and quantification of the bat white-nose syndrome fungus, Pseudogymnoascus destructans, from swab and environmental samples
Series title Journal of Veterinary Diagnostic Investigation
DOI 10.1177/1040638715626963
Volume 28
Issue 2
Year Published 2016
Language English
Publisher American Association of Veterinary Laboratory Diagnosticians
Publisher location Columbia, MO
Contributing office(s) National Wildlife Health Center
Description 9 p.
Larger Work Type Article
Larger Work Subtype Journal Article
Larger Work Title Journal of Veterinary Diagnostic Investigation
First page 110
Last page 118
Online Only (Y/N) N
Additional Online Files (Y/N) N