ArticleMicrobiome2021
Validation and standardization of DNA extraction and library construction methods for metagenomics-based human fecal microbiome measurements.
Article in Microbiome, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 57 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
57 citing papers in PubMed.
- Clinical benefits ofFrontiers in nutrition · 2025Trial
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- Oral care tablet containing kiwifruit powder affects tongue coating microbiome.Clinical and experimental dental research · 2022Trial
- Health Effects of Drinking Water Produced from Deep Sea Water: A Randomized Double-Blind Controlled Trial.Nutrients · 2022Trial
- Habitual Dietary Intake Affects the Altered Pattern of Gut Microbiome by Acarbose in Patients with Type 2 Diabetes.Nutrients · 2021Trial
- Comparative metagenomic assessment of Illumina-compatible library preparation methods, short-read lengths, and PacBio HiFi sequencing reveals differences in microbial and functional diversity recovery from a complex environmental sample.Microbiology spectrum · 2026Article
- Multi-omics profiling and bile-acid exposure assays implicate a gut microbiome-parasite axis linked to persistent Entamoeba histolytica carriage.Gut pathogens · 2026Article
- An Integrated Dataset of Clinical and Microbial Profiles for Fecal Microbiota Transplantation.Scientific data · 2026Article
- Analytic attributes of the 16S rRNA gene sequencing methodology for human gut microbiota characterization.Applied microbiology and biotechnology · 2026Article
- Profiling the Athletes' Gut Microbiome: A Critical Methodological Perspective on 16S Metabarcoding and Shotgun Metagenomics.Biology · 2026Review
- The microbiome landscape of oral cancer in young patients.JNCI cancer spectrum · 2026Article
- Investigation of Storage Conditions and Quality Control Markers for Metabolites and Lipids in Human Feces.Metabolites · 2026Article
- The gut microbiome in colorectal cancer: molecular paradigms and translational frontiers.Frontiers in bioengineering and biotechnology · 2026Review
- Gut microbiota and the early prevention window in type 1 diabetes and latent autoimmune diabetes in adults: a state-of-the-art narrative review on diet and metabolites.Frontiers in endocrinology · 2026Review
- The Japanese gut microbiome: ecology, uniqueness, and impact on health and disease.Proceedings of the Japan Academy. Series B, Physical and biological sciences · 2026Review
- Gut Microbiota in Hepatocellular Carcinoma: Etiology-Specific Evidence, Mechanisms, and Translational Challenges.Journal of hepatocellular carcinoma · 2026Review
- Genomic GC bias correction improves species abundance estimation from metagenomic data.Nature communications · 2025Article
Corrections and comments
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Authors and funding
35 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundValidation and standardization of methodologies for microbial community measurements by high-throughput sequencing are needed to support human microbiome research and its industrialization. This study set out to establish standards-based solutions to improve the accuracy and reproducibility of metagenomics-based microbiome profiling of human fecal samples.
resultsIn the first phase, we performed a head-to-head comparison of a wide range of protocols for DNA extraction and sequencing library construction using defined mock communities, to identify performant protocols and pinpoint sources of inaccuracy in quantification. In the second phase, we validated performant protocols with respect to their variability of measurement results within a single laboratory (that is, intermediate precision) as well as interlaboratory transferability and reproducibility through an industry-based collaborative study. We further ascertained the performance of our recommended protocols in the context of a community-wide interlaboratory study (that is, the MOSAIC Standards Challenge). Finally, we defined performance metrics to provide best practice guidance for improving measurement consistency across methods and laboratories.
conclusionsThe validated protocols and methodological guidance for DNA extraction and library construction provided in this study expand current best practices for metagenomic analyses of human fecal microbiota. Uptake of our protocols and guidelines will improve the accuracy and comparability of metagenomics-based studies of the human microbiome, thereby facilitating development and commercialization of human microbiome-based products. Video Abstract.
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