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The elusive resistome: a global comparison reveals large discrepancies among detection pipelines

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Title:The elusive resistome: a global comparison reveals large discrepancies among detection pipelines
Creators: Inda-Díaz, Juan S. ORCID logoORCID: https://orcid.org/0000-0002-3735-8300, Adegoke, Faith ORCID logoORCID: https://orcid.org/0000-0003-3835-590X, Löber, Ulrike ORCID logoORCID: https://orcid.org/0000-0001-7468-9531, Jarquín-Díaz, Víctor Hugo ORCID logoORCID: https://orcid.org/0000-0003-3758-1091, Duan, Yiqian ORCID logoORCID: https://orcid.org/0000-0002-8046-9831, Bengtsson-Palme, Johan ORCID logoORCID: https://orcid.org/0000-0002-6528-3158, Ugarcina Perovic, Svetlana ORCID logoORCID: https://orcid.org/0000-0001-7907-1597 and Coelho, Luis Pedro ORCID logoORCID: https://orcid.org/0000-0002-9280-7885
Abstract:Identifying antibiotic resistance genes (ARGs) from metagenomic data is critical for studying antimicrobial resistance across microbial communities and pathogens. However, there is no standardized methodology for ARG annotation. Here, we compare ten commonly used ARG detection pipelines by analysing over 270 million prokaryotic genes from the Global Microbial Gene Catalogue across 13 distinct habitats. We observed up to a 45-fold difference in the number of reported ARGs, with a mean Jaccard index of only 16% between pipelines. Pipeline selection profoundly impacted downstream biological interpretations, with drastic changes to estimates of ARG relative abundance and richness, to the characterization of pan- and core-resistomes, and to the class-level composition of the inferred resistome. ARG detection pipelines make different, defensible trade-offs, and no single approach should be treated as authoritative. Therefore, users should justify and communicate choices carefully, as our analyses show that, taken uncritically, the same data can support conflicting biological and ecological interpretations.
Keywords:Animals
Source:bioRxiv
Publisher:Cold Spring Harbor Laboratory Press
Article Number:2026.05.11.724158
Date:12 May 2026
Official Publication:https://doi.org/10.64898/2026.05.11.724158
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