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Evaluating the Information Content of Shallow Shotgun Metagenomics.

作者信息

Hillmann Benjamin, Al-Ghalith Gabriel A, Shields-Cutler Robin R, Zhu Qiyun, Gohl Daryl M, Beckman Kenneth B, Knight Rob, Knights Dan

机构信息

Department of Computer Science and Engineering, University of Minnesota, Minneapolis, Minnesota, USA.

Bioinformatics and Computational Biology, University of Minnesota, Minneapolis, Minnesota, USA.

出版信息

mSystems. 2018 Nov 13;3(6). doi: 10.1128/mSystems.00069-18. eCollection 2018 Nov-Dec.


DOI:10.1128/mSystems.00069-18
PMID:30443602
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6234283/
Abstract

Although microbial communities are associated with human, environmental, plant, and animal health, there exists no cost-effective method for precisely characterizing species and genes in such communities. While deep whole-metagenome shotgun (WMS) sequencing provides high taxonomic and functional resolution, it is often prohibitively expensive for large-scale studies. The prevailing alternative, 16S rRNA gene amplicon (16S) sequencing, often does not resolve taxonomy past the genus level and provides only moderately accurate predictions of the functional profile; thus, there is currently no widely accepted approach to affordable, high-resolution, taxonomic, and functional microbiome analysis. To address this technology gap, we evaluated the information content of shallow shotgun sequencing with as low as 0.5 million sequences per sample as an alternative to 16S sequencing for large human microbiome studies. We describe a library preparation protocol enabling shallow shotgun sequencing at approximately the same per-sample cost as 16S sequencing. We analyzed multiple real and simulated biological data sets, including two novel human stool samples with ultradeep sequencing of 2.5 billion sequences per sample, and found that shallow shotgun sequencing recovers more-accurate species-level taxonomic and functional profiles of the human microbiome than 16S sequencing. We discuss the inherent limitations of shallow shotgun sequencing and note that 16S sequencing remains a valuable and important method for taxonomic profiling of novel environments. Although deep WMS sequencing remains the gold standard for high-resolution microbiome analysis, we recommend that researchers consider shallow shotgun sequencing as a useful alternative to 16S sequencing for large-scale human microbiome research studies where WMS sequencing may be cost-prohibitive. A common refrain in recent microbiome-related academic meetings is that the field needs to move away from broad taxonomic surveys using 16S sequencing and toward more powerful longitudinal studies using shotgun sequencing. However, performing deep shotgun sequencing in large longitudinal studies remains prohibitively expensive for all but the most well-funded research labs and consortia, which leads many researchers to choose 16S sequencing for large studies, followed by deep shotgun sequencing on a subset of targeted samples. Here, we show that shallow- or moderate-depth shotgun sequencing may be used by researchers to obtain species-level taxonomic and functional data at approximately the same cost as amplicon sequencing. While shallow shotgun sequencing is not intended to replace deep shotgun sequencing for strain-level characterization, we recommend that microbiome scientists consider using shallow shotgun sequencing instead of 16S sequencing for large-scale human microbiome studies.

摘要
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0be2/6234283/ab0c2b3d9301/sys0051822770004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0be2/6234283/7d09fec086a7/sys0051822770001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0be2/6234283/0afe9d65d231/sys0051822770002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0be2/6234283/03aeaf91e9e8/sys0051822770003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0be2/6234283/ab0c2b3d9301/sys0051822770004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0be2/6234283/7d09fec086a7/sys0051822770001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0be2/6234283/0afe9d65d231/sys0051822770002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0be2/6234283/03aeaf91e9e8/sys0051822770003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0be2/6234283/ab0c2b3d9301/sys0051822770004.jpg

相似文献

[1]
Evaluating the Information Content of Shallow Shotgun Metagenomics.

mSystems. 2018-11-13

[2]
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[3]
Shallow shotgun sequencing reduces technical variation in microbiome analysis.

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[4]
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[5]
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[6]
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[7]
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引用本文的文献

[1]
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[2]
Integrative analysis of microbial 16S gene and shotgun metagenomic sequencing data improves statistical efficiency in testing differential abundance.

J Am Stat Assoc. 2025-8-5

[3]
Eukaryotic composition across seasons and social groups in the gut microbiota of wild baboons.

Anim Microbiome. 2025-6-21

[4]
Tracing non-fungal eukaryotic diversity via shotgun metagenomes in the complex mudflat intertidal zones.

mSystems. 2025-7-22

[5]
Consistent microbial insights across sequencing methods in soil studies: the role of reference taxonomies.

mSystems. 2025-7-22

[6]
Analyzing human gut microbiome data from global populations: challenges and resources.

Trends Microbiol. 2025-6-6

[7]
DNA extraction protocols for animal fecal material on blood spot cards.

PLoS One. 2025-5-12

[8]
Faecal metagenomes of great tits and blue tits provide insights into host, diet, pathogens and microbial biodiversity.

Access Microbiol. 2025-4-28

[9]
Metagenomic source tracking after microbiota transplant therapy.

Gut Microbes. 2025-12

[10]
Long-read metagenomics gives a more accurate insight into the microbiota of long-ripened gouda cheeses.

Front Microbiol. 2025-3-24

本文引用的文献

[1]
Assembly and ecological function of the root microbiome across angiosperm plant species.

Proc Natl Acad Sci U S A. 2018-1-22

[2]
A communal catalogue reveals Earth's multiscale microbial diversity.

Nature. 2017-11-23

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Large-scale differences in microbial biodiversity discovery between 16S amplicon and shotgun sequencing.

Sci Rep. 2017-7-31

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Genome Res. 2016-12

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PLoS One. 2016-11-7

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Characterization of the Gut Microbiome Using 16S or Shotgun Metagenomics.

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Library preparation methodology can influence genomic and functional predictions in human microbiome research.

Proc Natl Acad Sci U S A. 2015-11-10

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Nat Rev Microbiol. 2014-9

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