Negativeome characterization and decontamination in early-life virome studies

Abstract Contaminant sequences of external origin complicate the study of host-associated viromes, particularly in low-biomass samples obtained through viral-like particle (VLP) enrichment. However, the prevalence and impact of external contaminants on low-biomass samples are under-studied. Here, we...

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Main Authors: Nataliia Kuzub, Alexander Kurilshikov, Alexandra Zhernakova, Sanzhima Garmaeva
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-61478-7
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author Nataliia Kuzub
Alexander Kurilshikov
Alexandra Zhernakova
Sanzhima Garmaeva
author_facet Nataliia Kuzub
Alexander Kurilshikov
Alexandra Zhernakova
Sanzhima Garmaeva
author_sort Nataliia Kuzub
collection DOAJ
description Abstract Contaminant sequences of external origin complicate the study of host-associated viromes, particularly in low-biomass samples obtained through viral-like particle (VLP) enrichment. However, the prevalence and impact of external contaminants on low-biomass samples are under-studied. Here, we analyze 1321 gut virome samples and 55 negative controls (NCs) from four early-life virome studies. Virus sequences identified in NCs, termed negativeome, were used as a proxy for the contamination assessment. We show that 61% of samples share at least one identical strain with negativeome, likely representing external contamination. While the median abundance of contaminant strains in these samples is only 1%, it ranges from 0 to 99% and exceeds 10% in 11% of infant samples. We further demonstrate that contamination is largely study-specific and has a greater impact on infant samples than on maternal samples. Based on our results, we propose a contamination assessment method using a publicly available database of sequences detected in NCs and a strain-level decontamination strategy.
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spelling doaj-art-a4cb457fd307400a902a4031f23355702025-08-20T03:45:33ZengNature PortfolioNature Communications2041-17232025-07-0116111210.1038/s41467-025-61478-7Negativeome characterization and decontamination in early-life virome studiesNataliia Kuzub0Alexander Kurilshikov1Alexandra Zhernakova2Sanzhima Garmaeva3Department of Genetics, University of Groningen, University Medical Center GroningenDepartment of Genetics, University of Groningen, University Medical Center GroningenDepartment of Genetics, University of Groningen, University Medical Center GroningenDepartment of Genetics, University of Groningen, University Medical Center GroningenAbstract Contaminant sequences of external origin complicate the study of host-associated viromes, particularly in low-biomass samples obtained through viral-like particle (VLP) enrichment. However, the prevalence and impact of external contaminants on low-biomass samples are under-studied. Here, we analyze 1321 gut virome samples and 55 negative controls (NCs) from four early-life virome studies. Virus sequences identified in NCs, termed negativeome, were used as a proxy for the contamination assessment. We show that 61% of samples share at least one identical strain with negativeome, likely representing external contamination. While the median abundance of contaminant strains in these samples is only 1%, it ranges from 0 to 99% and exceeds 10% in 11% of infant samples. We further demonstrate that contamination is largely study-specific and has a greater impact on infant samples than on maternal samples. Based on our results, we propose a contamination assessment method using a publicly available database of sequences detected in NCs and a strain-level decontamination strategy.https://doi.org/10.1038/s41467-025-61478-7
spellingShingle Nataliia Kuzub
Alexander Kurilshikov
Alexandra Zhernakova
Sanzhima Garmaeva
Negativeome characterization and decontamination in early-life virome studies
Nature Communications
title Negativeome characterization and decontamination in early-life virome studies
title_full Negativeome characterization and decontamination in early-life virome studies
title_fullStr Negativeome characterization and decontamination in early-life virome studies
title_full_unstemmed Negativeome characterization and decontamination in early-life virome studies
title_short Negativeome characterization and decontamination in early-life virome studies
title_sort negativeome characterization and decontamination in early life virome studies
url https://doi.org/10.1038/s41467-025-61478-7
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AT sanzhimagarmaeva negativeomecharacterizationanddecontaminationinearlylifeviromestudies