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Performance criteria for the analyses of environmental DNA by targeted quantitative polymerase chain reaction
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This is the first edition of CSA W219, Performance criteria for the analyses of environmental DNA by targeted quantitative polymerase chain reaction. It is a companion Standard to CSA W214.
Users of this Standard should note that additional eDNA study design, methodology, and reporting requirements might be specified by federal, provincial/territorial, municipal, or other authorities, or by a project owner, depending on the larger study objectives. This Standard should not be considered as a replacement for the requirements contained in any
a) applicable federal, territorial, or provincial statute;
b) regulation, licence, or permit issued pursuant to an applicable statute; or
c) contract that an owner has with a contractor.
CSA Group acknowledges that the development of this Standard was made possible by the financial support of Genome Canada through the large scale applied research project program.
This Standard was prepared by the Technical Committee on Targeted qPCR-Based eDNA Assays, under the jurisdiction of the Strategic Steering Committee on Natural Resources, and has been formally approved by the Technical Committee.
This Standard has been developed in compliance with Standards Council of Canada requirements for National Standards of Canada. It has been published as a National Standard of Canada by CSA Group.
This Standard applies to targeted qPCR-based eDNA assays used to conduct eDNA biological surveys or assessments.
This Standard provides performance criteria pertaining to
b) analytical performance including
iv) repeatability; and
v) reproducibility; and
c) field application criteria including
i) verification of target taxon detection using a sample matrix;
ii) technical replicates run per environmental sample;
iii) assay controls for false positive (Type 1) errors; and
iv) assay controls for false negative (Type 2) errors.
This Standard does not address field components, including sampling strategy and methodology, or interpretation of field survey results that accompany data generated from targeted eDNA assays.
Users should be aware the requirements in this document might be influenced by those in future Standards that will address other steps (e.g., sampling design, sampling methods) in an eDNA assay.
Note: Supporting technical background information and rationales are provided in Annex A.
This Standard is intended for use by testing entities (including commercial, industrial, government, and academic laboratories) developing and using eDNA assays.
The Standard can also be used by
a) industrial and commercial proponents of resource development projects whose businesses can have direct or indirect influences on aquatic and other ecosystems;
b) regulatory agencies including but not limited to federal, Indigenous, provincial, and municipal resource managers;
c) contract administrators who oversee projects that use any aspect of eDNA methodology;
d) non-government stakeholders, including environmental non-government organizations, that might undertake eDNA monitoring studies or review/audit eDNA projects for compliance with reasonable expectations for due diligence;
e) qualified environmental professionals who undertake eDNA monitoring studies or audit projects to ensure competency and completeness;
f) academic researchers and educators, for the purposes of improving and advancing the collective field of genetics research; and
g) others with a vested interest in aspects of eDNA studies including project design, execution, and results interpretation.
The requirements provided in this Standard apply to practitioners engaged in the delivery of targeted qPCR-based eDNA studies and the reporting of their results. Project results could influence decisions affecting the conservation and management of natural resources. The intent of this Standard is that its application enables effective evaluation of methods during project implementation and enhances confidence in decision-making based on the project results.
1.4 Intended use
This Standard applies to the performance criteria of targeted qPCR assays used in eDNA studies and surveys. Used in conjunction with CSA W214, this Standard should also be used in conjunction with other Standards (e.g., ISO/IEC 17025) to enable laboratories to demonstrate they operate competently and generate valid results.
While this Standard focuses on the most widely used eDNA detection methods, it is recognized that other methods might also be suitable. This Standard is not intended to constrain future developments in eDNA approaches, methodologies, or implementations.
The elements of this Standard are relevant to multiple steps in the eDNA workflow addressed in CSA W214. Users are advised to refer to both this Standard and CSA W214 in their entirety.
Where quantitative measurements are required, users should report these measurements using relevant standard measures and units.
In this Standard, shall is used to express a requirement, i.e., a provision that the user is obliged to satisfy to comply with the Standard; should is used to express a recommendation or that which is advised but not required; and may is used to express an option or that which is permissible within the limits of the Standard.
Notes accompanying clauses do not include requirements or alternative requirements; the purpose of a note accompanying a clause is to separate explanatory or informative material from the text.
Notes to tables and figures are considered part of the table or figure and may be written as requirements.
Annexes are designated normative (mandatory) or informative (non-mandatory) to define their application.
Environmental DNA (eDNA) reporting requirements and terminology