This document provides guidance on the detection and quantification of nucleic acids (NA) from environmental samples using polymerase chain reaction (PCR). Environmental Nucleic Acids (eNA) include environmental DNA (eDNA) and environmental RNA (eRNA). This document addresses the validation and application of PCR-based assays, with a focus on macroorganisms. The detection of nucleic acids from microbial and viral sources is discussed in separate documents (ISO/TS 12896; ISO 16099). Specifically, this standard is designed for PCR-based eDNA/eRNA assays for the detection of specific, targeted taxa, and not for community-wide or assemblage detection using metabarcoding.
This standard covers eNA amplification from environmental samples of varying matrices, including but not limited to water, air, soil/sediment, fecal samples, and bulk samples. Methods described include quantitative PCR (qPCR), digital PCR (dPCR), and reverse-transcription PCR for species-targeted detection. It also describes the methodological controls required when developing an assay and when using an assay with eNA samples. Finally, this document includes reporting requirements for assay development and assay use.
This document excludes procedures that occur before PCR (e.g., eDNA extractions) and after PCR (e.g., interpreting results in the context of a study’s objectives).
Registration number (WIID)
93817
Scope
This document provides guidance on the detection and quantification of nucleic acids (NA) from environmental samples using polymerase chain reaction (PCR). Environmental Nucleic Acids (eNA) include environmental DNA (eDNA) and environmental RNA (eRNA). This document addresses the validation and application of PCR-based assays, with a focus on macroorganisms. The detection of nucleic acids from microbial and viral sources is discussed in separate documents (ISO/TS 12896; ISO 16099). Specifically, this standard is designed for PCR-based eDNA/eRNA assays for the detection of specific, targeted taxa, and not for community-wide or assemblage detection using metabarcoding.
This standard covers eNA amplification from environmental samples of varying matrices, including but not limited to water, air, soil/sediment, fecal samples, and bulk samples. Methods described include quantitative PCR (qPCR), digital PCR (dPCR), and reverse-transcription PCR for species-targeted detection. It also describes the methodological controls required when developing an assay and when using an assay with eNA samples. Finally, this document includes reporting requirements for assay development and assay use.
This document excludes procedures that occur before PCR (e.g., eDNA extractions) and after PCR (e.g., interpreting results in the context of a study’s objectives).
