# A QIIME2-based workflow for multi-amplicon 16S rRNA profiling

**Authors:** Armando G. Licata, Marica Zoppi, Chiara Dossena, Federico Rossignoli, Davide Rizzo, Luca Bergamaschi, Olga Nigro, Stefano Chiaravalli, Maura Massimino, Loris De Cecco

PMC · DOI: 10.1128/mra.00836-25 · 2025-12-09

## TL;DR

This paper introduces a QIIME2-based workflow for analyzing 16S rRNA sequencing data from multiple amplicon regions.

## Contribution

The novel contribution is a validated open-source pipeline that matches proprietary tools in accuracy and depth for multi-amplicon 16S rRNA sequencing.

## Key findings

- The workflow achieves a taxonomic F1-score of 0.875 when benchmarked against a mock community.
- Multi-region sequencing outperforms single amplicon approaches in taxonomic resolution.
- The pipeline is robust for semiconductor-based sequencing data.

## Abstract

We present an open-source QIIME2 pipeline for 16S multi-amplicon sequencing. Benchmarked against proprietary software with a mock community, our workflow demonstrates comparable sequencing depth and taxonomic accuracy (F1-Score=0.875). The multi-region approach outperforms single amplicons, validating our pipeline as a robust alternative for semiconductor-based sequencing data.

## Full-text entities

- **Chemicals:** MAN0010799 (-)
- **Species:** Escherichia coli (E. coli, species) [taxon 562]

## Figures

1 figure with captions in the complete paper: https://tomesphere.com/paper/PMC12797943/full.md

---
Source: https://tomesphere.com/paper/PMC12797943