title: REPRODUCIBLE RESEARCH WORKFLOW IN R FOR THE ANALYSIS OF PERSONALIZED HUMAN MICROBIOME DATA.
abstract: This article presents a reproducible research workflow for amplicon-based microbiome studies in personalized medicine created using Bioconductor packages and the knitr markdown interface.We show that sometimes a multiplicity of choices and lack of consistent documentation at each stage of the sequential processing pipeline used for the analysis of microbiome data can lead to spurious results. We propose its replacement with reproducible and documented analysis using R packages dada2, knitr, and phyloseq. This workflow implements both key stages of amplicon analysis: the initial filtering and denoising steps needed to construct taxonomic feature tables from error-containing sequencing reads (dada2), and the exploratory and inferential analysis of those feature tables and associated sample metadata (phyloseq). This workow facilitates reproducible interrogation of the full set of choices required in microbiome studies. We present several examples in which we leverage existing packages for analysis in a way that allows easy sharing and modification by others, and give pointers to articles that depend on this reproducible workflow for the study of longitudinal and spatial series analyses of the vaginal microbiome in pregnancy and the oral microbiome in humans with healthy dentition and intra-oral tissues.
authors.affiliation: Statistics Department, Stanford, CA 94305, USA.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Callahan
authors.forename: Diana
authors.initials: D
authors.lastname: Proctor
authors.forename: David
authors.initials: D
authors.lastname: Relman
authors.forename: Julia
authors.initials: J
authors.lastname: Fukuyama
authors.forename: Susan
authors.initials: S
authors.lastname: Holmes
mesh_terms: D019295:Computational Biology / Q000379:methods / Q000706:statistics & numerical data; D003627:Data Interpretation, Statistical; D005260:Female; D059014:High-Throughput Nucleotide Sequencing / Q000706:statistics & numerical data; D006801:Humans; D008137:Longitudinal Studies; D059013:Microbial Consortia / Q000235:genetics; D064307:Microbiota / Q000235:genetics; D009055:Mouth / Q000382:microbiology; D057285:Precision Medicine / Q000706:statistics & numerical data*; D011247:Pregnancy; D025341:Principal Component Analysis; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA / Q000706:statistics & numerical data; D012984:Software; D014621:Vagina / Q000382:microbiology; D057188:Workflow