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White-Nose Syndrome Diagnostic Laboratory Network Handbook

Techniques and Methods 15-E1
By:  and 
https://doi.org/10.3133/tm15E1

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Abstract

When responding to a wildlife disease outbreak, managers depend on consistent and clear data to make decisions. However, diagnostic methods for detecting pathogens of wildlife often lack the level of procedural and interpretational standardization that occurs in the investigation of human and domestic animal diseases. This lack of standardization can hamper diagnostic reliability in two ways. First is the inappropriate application of tests to new species or in situations that are outside of the original (in other words, validated) purpose. Second is the use of laboratory-specific modifications or analytical parameters without thorough investigation of how those changes affect result comparisons across institutions or the ability to make broader conclusions about pathogen or disease.

White-nose syndrome (WNS) is a disease caused by the fungal pathogen Pseudogymnoascus destructans (Pd), which has spread rapidly and is causing population-level declines in some species of North American bats. During the last decade, quantitative polymerase chain reaction (qPCR) has become the most common method of testing for Pd because of qPCR’s speed, accuracy, and simplicity across a wide range of invasive and noninvasive sample types. Its widespread use by many State, Federal, Provincial, and academic institutions has inevitably led to variations in methodology and interpretation among laboratories. The progressive geographic spread of fungus and disease has also led to sampling contexts and strategies that differ from those for which the qPCR assay was originally developed and validated. These factors have resulted in inconsistencies among results tested in different laboratories and, subsequently, confusion for managers and decision makers.

To address these challenges, the WNS National Response Team Diagnostic Working Group launched a project congruent with increased calls for the harmonization of wildlife disease diagnostic results, and reporting standards across disparate methodologies and laboratories. Beginning in 2019, interlaboratory testing was done to better understand how variations to Pd qPCR methodology affect diagnostic consistency and to reassess the assay’s fit for purpose in new testing contexts. This information led to expanded conversations within the Diagnostic Working Group related to best practices in Pd qPCR diagnostic testing, the development of common interpretation language for classifying test results, and the incorporation of that language into an updated WNS case definition. This handbook is the resulting product and is intended to help further harmonize Pd qPCR diagnostic testing by establishing recommendations related to voluntary participation in a WNS Diagnostic Laboratory Network, documenting the currently (2022) practiced Pd qPCR methodologies, discussing general best practices for molecular diagnostics and laboratory networks, and elaborating on the epidemiologic and diagnostic basis of the agreed-upon classification language for Pd qPCR results. Through this voluntary, consensus-based approach to diagnostic harmonization, this work aims to improve the confidence of management agencies in reported Pd qPCR results and can serve as an example of national diagnostic coordination for other unregulated wildlife diseases.

Suggested Citation

Alger, K., and White Nose Syndrome National Response Team Diagnostic Working Group, 2023, White-Nose Syndrome Diagnostic Laboratory Network handbook: U.S. Geological Survey Techniques and Methods, book 15, chap. E1, 50 p., https://doi.org/10.3133/tm15E1.

ISSN: 2328-7055 (online)

Table of Contents

  • Acknowledgments
  • Abstract
  • Introduction
  • White-Nose Syndrome Response Team Diagnostic and Surveillance Working Group
  • Standardization Versus Harmonization
  • Purpose and Scope
  • Principles of Wildlife Disease Sampling with Additional Resources
  • Sampling Considerations for Pseudogymnoascus destructans
  • Laboratory Biosecurity and Quality Management Systems
  • Pseudogymnoascus destructans Molecular Detection Methods (Deoxyribonucleic Acid Extraction and Quantitative Polymerase Chain Reaction)
  • Best Management Practices for Laboratory Network Participation
  • Summary
  • References Cited
Publication type Report
Publication Subtype USGS Numbered Series
Title White-Nose Syndrome Diagnostic Laboratory Network handbook
Series title Techniques and Methods
Series number 15-E1
DOI 10.3133/tm15E1
Year Published 2023
Language English
Publisher U.S. Geological Survey
Publisher location Reston, VA
Contributing office(s) National Wildlife Health Center
Description Report: x, 50 p.; Data Release
Online Only (Y/N) Y
Google Analytic Metrics Metrics page
Additional publication details