Abstract

With this review, we aim to increase the quality standards for clinical studies with microbiome as an output parameter. We critically address the existing body of evidence for good quality practices in oral microbiome studies based on 16S rRNA gene amplicon sequencing. First, we discuss the usefulness of microbiome profile analyses. Is a microbiome study actually the best approach for answering the research question? This is followed by addressing the criteria for the most appropriate study design, sample size, and the necessary data (study metadata) that should be collected. Next, we evaluate the available evidence for best practices in sample collection, transport, storage, and DNA isolation. Finally, an overview of possible sequencing options (eg, 16S rRNA gene hypervariable regions, sequencing platforms), processing and data interpretation approaches, as well as requirements for meaningful data storage, sharing, and reporting are provided.

Highlights

  • About a decade ago, when the first publications on the oral microbiome using high throughput 16S ribosomal RNA gene amplicon sequencing appeared,[1,2,3,4] the methodologies of sample processing, sequencing, and the downstream bioinformatic analyses still had to evolve

  • Several studies report a relation between income or education level and oral microbial composition: a microarray study on stimulated saliva of 292 Danish adults with low levels of caries and periodontitis found that socioeconomic status explained 20% of the overall variance in salivary microbiome.[29]

  • A relevant question regarding sample choice in periodontitis patients is if one has to collect subgingival plaque in order to assess the effects of interventions

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Summary

Link to publication

Citation for published version (APA): Zaura, E., Pappalardo, V. Disclaimer/Complaints regulations If you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: https://uba.uva.nl/en/contact, or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. UvA-DARE is a service provided by the library of the University of Amsterdam (https://dare.uva.nl)

REVIEW ARTICLE
| INTRODUCTION
Metadata for samples in oral microbiome studies
Positive controls
OMNIgene saliva kit
Scope mouthwash
Main results
Illumina MiSeq
Cell lysis with ZB and L more effective than other methods
High error rate
Findings
Quality control of raw sequencing data
Full Text
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