Title

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

Files

Link to Full Text

Download Full Text

Publication Date

March 2016

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.

Keywords

Bats, DNA Extraction, Pseudogymnoascus Destructans, Quantification, White-Nose Syndrome.

Description

RDA

Subject: topical

Bats; DNA Extraction; Pseudogymnoascus Destructans; Quantification; White-Nose Syndrome.

Type

Article

Genre

serial

Identifier

SFS0044367_00001

Share

COinS