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      Multi-group Analysis of Compositions of Microbiomes with Covariate Adjustments and Repeated Measures

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      American Journal Experts

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          Abstract

          Microbiome differential abundance analysis methods for a pair of groups are well established in the literature. However, many microbiome studies involve multiple groups, sometimes even ordered groups, such as stages of a disease, and require different types of comparisons. Standard pairwise comparisons are not only inefficient in terms of power and false discovery rates, but they may not address the scientific question of interest. In this paper, we propose a general framework for performing a wide range of multi-group analyses with covariate adjustments and repeated measures. We demonstrate the effectiveness of our methodology through two real data sets. The first example explores the effects of aridity on the soil microbiome, and the second example investigates the effects of surgical interventions on the microbiome of IBD patients.

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          Most cited references54

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          Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing

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            Analysis of composition of microbiomes: a novel method for studying microbial composition

            Background Understanding the factors regulating our microbiota is important but requires appropriate statistical methodology. When comparing two or more populations most existing approaches either discount the underlying compositional structure in the microbiome data or use probability models such as the multinomial and Dirichlet-multinomial distributions, which may impose a correlation structure not suitable for microbiome data. Objective To develop a methodology that accounts for compositional constraints to reduce false discoveries in detecting differentially abundant taxa at an ecosystem level, while maintaining high statistical power. Methods We introduced a novel statistical framework called analysis of composition of microbiomes (ANCOM). ANCOM accounts for the underlying structure in the data and can be used for comparing the composition of microbiomes in two or more populations. ANCOM makes no distributional assumptions and can be implemented in a linear model framework to adjust for covariates as well as model longitudinal data. ANCOM also scales well to compare samples involving thousands of taxa. Results We compared the performance of ANCOM to the standard t-test and a recently published methodology called Zero Inflated Gaussian (ZIG) methodology (1) for drawing inferences on the mean taxa abundance in two or more populations. ANCOM controlled the false discovery rate (FDR) at the desired nominal level while also improving power, whereas the t-test and ZIG had inflated FDRs, in some instances as high as 68% for the t-test and 60% for ZIG. We illustrate the performance of ANCOM using two publicly available microbial datasets in the human gut, demonstrating its general applicability to testing hypotheses about compositional differences in microbial communities. Conclusion Accounting for compositionality using log-ratio analysis results in significantly improved inference in microbiota survey data.
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              Linear Statistical Inference and its Applications

              C. Rao (1973)
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                Author and article information

                Journal
                Res Sq
                ResearchSquare
                Research Square
                American Journal Experts
                02 May 2023
                : rs.3.rs-2778207
                Affiliations
                [1 ]Biostatistics and Computational Biology Branch, NIEHS, NIH, Research Triangle Park, NC, USA
                Author notes

                Author contributions

                Both authors contributed equally to the theory and methodology described in this paper. All numerical works and computations were conducted by H.L. who developed ANCOM-BC2 pipeline in R that is freely and publicly available. Please contact H.L. for software requests.

                Author information
                http://orcid.org/0000-0003-1276-7252
                Article
                10.21203/rs.3.rs-2778207
                10.21203/rs.3.rs-2778207/v1
                10187376
                37205444
                076f4c15-0656-4510-bdd4-11aecb92f84e

                This work is licensed under a Creative Commons Attribution 4.0 International License, which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use.

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