Overview - ISO 20397-3:2025 (Metagenomics, MPS)
ISO 20397-3:2025 is the first-edition International Standard that defines general requirements and guidance for metagenomics workflows using massively parallel sequencing (MPS). Intended for laboratories and research organizations, the standard covers end-to-end processes for metagenomics from primary sample handling through sequencing, bioinformatics analysis, and validation. It is Part 3 of the ISO 20397 series and complements Parts 1 and 2 on nucleic acid/library preparation and sequencing data quality.
Key SEO keywords: metagenomics standard, massively parallel sequencing (MPS), bioinformatics pipeline, library preparation, sample preparation.
Key Topics and Technical Requirements
The document specifies structured requirements across the complete metagenomics pipeline:
- Sampling strategy and sample handling
- Primary sample types, stabilization, storage, transportation and extraction considerations
- DNA/RNA isolation and nucleic acid sample quality assessment before library construction
- Nucleic acid library preparation
- Library construction practices for DNA/cDNA/RNA suited to MPS platforms
- Design and sequencing strategy
- Guidelines for short-read, long-read and hybrid sequencing approaches
- Sample preparation, sequencing assessment and design review processes
- Database construction
- Guidance for using public reference databases and building/curating self-built databases for taxonomic and functional annotation
- Bioinformatics analysis and reporting
- Requirements for bioinformatics pipelines, sequence alignment, taxonomic identification lists, functional annotation, and report content
- Validation and verification
- Pipeline and database validation using in silico controls and real sample controls; performance evaluation, verification and documentation
- Reporting and evaluation
- Test report content and evaluation criteria for metagenomics outputs
Normative references include ISO 20397-1, ISO 20397-2, ISO/TS 24420 and ISO 20395.
Practical Applications and Users
ISO 20397-3 is useful for organizations performing metagenomics in:
- Environmental and ecological monitoring (microbial community profiling)
- Clinical microbiology and infectious disease research
- Agricultural and food-safety microbiome studies
- Public health surveillance and biodefense
- Core sequencing facilities and bioinformatics service providers
Primary users: laboratory managers, quality managers, molecular biologists, sequencing technicians, bioinformaticians, database curators and auditors.
Related Standards
- ISO 20397-1: Nucleic acid and library preparation
- ISO 20397-2: Quality evaluation of sequencing data
- ISO/TS 24420: Data processing for shotgun metagenomics
- ISO 20395: Performance evaluation of nucleic acid quantification methods
ISO 20397-3 provides a harmonized framework for robust, reproducible metagenomics workflows and for validating bioinformatics pipelines used in MPS-based microbial community analysis.
Open preview (PDF)Biotechnology — Massively parallel sequencing — Part 3: General requirements and guidance for metagenomics
