Microbiome differential abundance methods produce different results across 38 datasets

Jacob T. Nearing (), Gavin M. Douglas, Molly G. Hayes, Jocelyn MacDonald, Dhwani K. Desai, Nicole Allward, Casey M. A. Jones, Robyn J. Wright, Akhilesh S. Dhanani, André M. Comeau and Morgan G. I. Langille
Additional contact information
Jacob T. Nearing: Dalhousie University
Gavin M. Douglas: Dalhousie University
Molly G. Hayes: Dalhousie University
Jocelyn MacDonald: Dalhousie University
Dhwani K. Desai: Dalhousie University
Nicole Allward: Dalhousie University
Casey M. A. Jones: Dalhousie University
Robyn J. Wright: Dalhousie University
Akhilesh S. Dhanani: Dalhousie University
André M. Comeau: Dalhousie University
Morgan G. I. Langille: Dalhousie University

Nature Communications, 2022, vol. 13, issue 1, 1-16

Abstract: Abstract Identifying differentially abundant microbes is a common goal of microbiome studies. Multiple methods are used interchangeably for this purpose in the literature. Yet, there are few large-scale studies systematically exploring the appropriateness of using these tools interchangeably, and the scale and significance of the differences between them. Here, we compare the performance of 14 differential abundance testing methods on 38 16S rRNA gene datasets with two sample groups. We test for differences in amplicon sequence variants and operational taxonomic units (ASVs) between these groups. Our findings confirm that these tools identified drastically different numbers and sets of significant ASVs, and that results depend on data pre-processing. For many tools the number of features identified correlate with aspects of the data, such as sample size, sequencing depth, and effect size of community differences. ALDEx2 and ANCOM-II produce the most consistent results across studies and agree best with the intersect of results from different approaches. Nevertheless, we recommend that researchers should use a consensus approach based on multiple differential abundance methods to help ensure robust biological interpretations.

Date: 2022
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DOI: 10.1038/s41467-022-28034-z

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