Evidence map›Paper›PMID 42919172›Full record

ArticleBMC microbiology2026

Evaluating 16S metagenomics data analysis pipelines using an Ion Torrent sequencer for human gut microbiome research.

Roddy Jorquera, Troy Ejsmentewicz, Rodrigo Lagos, Nicolas Bravo, Francisco Issotta, Andrés Hernández-Oliveras, Franz Villarroel-Espíndola, Roxana González-Stegmaier

Abstract readEvaluation Study
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In one paragraph

Article in BMC microbiology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it
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0citing papers in PubMed
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1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors.

Roddy Jorquera *Fundación Arturo Lopez Perez OECI Cancer Center, Translational Medicine Laboratory, Centro de Investigación e Innovación en Cáncer, Santiago, Chile. roddy.jorquera@gmail.com.
Troy Ejsmentewicz *Fundación Arturo Lopez Perez OECI Cancer Center, Translational Medicine Laboratory, Centro de Investigación e Innovación en Cáncer, Santiago, Chile.
Rodrigo LagosFundación Arturo Lopez Perez OECI Cancer Center, Epidemiology and Clinical Research Unit, Centro de Investigación e Innovación en Cáncer, Santiago, Chile.
Nicolas BravoFundación Arturo Lopez Perez OECI Cancer Center, Informatic Medicine and Data Science Unit, Santiago, Chile.
Francisco IssottaCentro Científico y Tecnológico de Excelencia Ciencia & Vida, Santiago, Chile.
Andrés Hernández-OliverasFundación Arturo Lopez Perez OECI Cancer Center, Translational Medicine Laboratory, Centro de Investigación e Innovación en Cáncer, Santiago, Chile.
Franz Villarroel-EspíndolaFundación Arturo Lopez Perez OECI Cancer Center, Translational Medicine Laboratory, Centro de Investigación e Innovación en Cáncer, Santiago, Chile. franz.villarroel@falp.org.
Roxana González-StegmaierFundación Arturo Lopez Perez OECI Cancer Center, Translational Medicine Laboratory, Centro de Investigación e Innovación en Cáncer, Santiago, Chile. roxana.gonzalez@falp.org.

Funding

Fundacion Arturo Lopez Perez internal grant ID FALP-LMT-2021Fundacion Arturo Lopez Perez internal grant ID FALP-LMT-2022
6 · The paper itself

Abstract

backgroundHigh-throughput 16S rRNA gene amplicon sequencing is essential for characterizing gut dysbiosis in translational research. However, microbiome profiles vary widely across sequencing platforms, library preparation protocols, targeted hypervariable regions (HVRs), and bioinformatics workflows. For users of the Ion Torrent S5 platform, there is currently no consensus methodology for performing open-source analyses as an alternative to proprietary frameworks. This study evaluates the proprietary Ion Reporter™ software against eight open-source pipelines based on QIIME2 and the SILVA database, accounting for data compositionality. Benchmarking was performed using a commercial human fecal reference material (ZymoBIOMICS™ D6323) characterized via DNA-shotgun sequencing (the ZB-Shotgun profile), as the reference standard.

resultsPoisson regression modeling showed that using two primer sets for library preparation yielded a 30 % lower expected read count than a single primer set (IRR = 0.6986, p < 0.001). Across all configurations, the choice of denoising algorithm (DADA2 vs. Deblur) significantly affected alpha diversity calculations. At the phylum level, hierarchical clustering of Bayes factors (BFs) indicated that multi-region Ion Reporter workflows produced stable consensus predictions. Conversely, at the species level, single-HVR open-source pipelines significantly outperformed multi-region approaches.

conclusionsOpen-source pipelines that combine targeted HVR2 or HVR4 inputs with state-of-the-art denoisers outperform proprietary Ion Reporter options, providing superior taxonomic resolution and precision for species-level microbiome profiling. This study establishes a traceable, reproducible, and cost-effective framework that empowers independent researchers to maximize the utility of Ion Torrent 16S rRNA data in healthcare and gastrointestinal research without relying on restrictive software ecosystems.

Indexed as

BacteriaGastrointestinal MicrobiomeHigh-Throughput Nucleotide SequencingMetagenomicsRNA, Ribosomal, 16SComputational BiologyDNA, BacterialFecesHumansSequence Analysis, DNASoftwareDNA, BacterialRNA, Ribosomal, 16S16S rRNA gene profilingIon Torrent PGMNGS BiotechnologyTaxonomic resolutionTranslational research

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.