Reliability of species detection in 16S microbiome analysis: Comparison of five widely used pipelines and recommendations for a more standardized approach

• DOI: 10.1371/journal.pone.0280870
• Publication Date: 2023-02-16T18:39:04Z
• Authors: Andreas Hiergeist; Jean Ruelle; Stefan Emler; André Gessner
• Keywords: Ion semiconductor sequencing

The use of NGS-based testing the bacterial microbiota is often impeded by inconsistent or non-reproducible results, especially when applying different analysis pipelines and reference databases. We investigated five frequently used software packages submitting same monobacterial datasets to them, representing V1-2 V3-4 regions 16S-rRNA gene 26 well characterized strains, which were sequenced Ion Torrent™ GeneStudio S5 system. results obtained divergent calculations relative abundance did not yield expected 100%. these inconsistencies able attribute them failures either themselves databases they rely on. On basis findings, we recommend certain standards should help render microbiome more consistent reproducible, thus useful in clinical practice.