A metagenomic DNA sequencing assay that is robust against environmental DNA contamination

Nat Commun. 2022 Jul 21;13(1):4197. doi: 10.1038/s41467-022-31654-0.

Abstract

Metagenomic DNA sequencing is a powerful tool to characterize microbial communities but is sensitive to environmental DNA contamination, in particular when applied to samples with low microbial biomass. Here, we present Sample-Intrinsic microbial DNA Found by Tagging and sequencing (SIFT-seq) a metagenomic sequencing assay that is robust against environmental DNA contamination introduced during sample preparation. The core idea of SIFT-seq is to tag the DNA in the sample prior to DNA isolation and library preparation with a label that can be recorded by DNA sequencing. Any contaminating DNA that is introduced in the sample after tagging can then be bioinformatically identified and removed. We applied SIFT-seq to screen for infections from microorganisms with low burden in blood and urine, to identify COVID-19 co-infection, to characterize the urinary microbiome, and to identify microbial DNA signatures of sepsis and inflammatory bowel disease in blood.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • COVID-19*
  • DNA
  • DNA Contamination
  • DNA, Bacterial / genetics
  • DNA, Environmental*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Metagenomics
  • Sequence Analysis, DNA

Substances

  • DNA, Bacterial
  • DNA, Environmental
  • DNA