The relationships among bacterial flora, diseases, and diet have been described by many authors. An operational taxonomic units (OTUs) are the result of clustering the 16S rRNA gene sequences at a certain cutoff value, and they are considered compositional data. As Pearson’s correlation coefficient is difficult to interpret, Aitchison’s ratio analysis was used to develop a method to handle compositional data. Multivariate analysis was developed because univariate analysis can be subject to large biases. Simulations regarding absolute abundance based on certain assumptions and some analyses, such as nonparametric multidimensional scaling (NMDS), principal component analysis (PCA), and ratio analysis, were conducted in this study. The same content as a 100% stacked bar graph could be expressed in low dimensions using PCA. However, the relative diversity was not reproducible with NMDS. Various assumptions were made regarding absolute abundance based on the relative abundance. However, which assumptions are true could not be determined. In summary, ratio analysis and PCA are useful for analyzing compositional data and the gut microbiota.
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