Turning the needle into the haystack: Culture-independent amplification of complex microbial genomes directly from their native environment

PLoS Pathog. 2024 Sep 12;20(9):e1012418. doi: 10.1371/journal.ppat.1012418. eCollection 2024 Sep.

Abstract

High-throughput sequencing (HTS) has revolutionized microbiology, but many microbes exist at low abundance in their natural environment and/or are difficult, if not impossible, to culture in the laboratory. This makes it challenging to use HTS to study the genomes of many important microbes and pathogens. In this review, we discuss the development and application of selective whole genome amplification (SWGA) to allow whole or partial genomes to be sequenced for low abundance microbes directly from complex biological samples. We highlight ways in which genomic data generated by SWGA have been used to elucidate the population dynamics of important human pathogens and monitor development of antimicrobial resistance and the emergence of potential outbreaks. We also describe the limitations of this method and propose some potential innovations that could be used to improve the quality of SWGA and lower the barriers to using this method across a wider range of infectious pathogens.

Publication types

  • Review

MeSH terms

  • Bacteria / genetics
  • Genome, Bacterial
  • Genome, Microbial
  • High-Throughput Nucleotide Sequencing* / methods
  • Humans
  • Nucleic Acid Amplification Techniques / methods
  • Whole Genome Sequencing / methods

Grants and funding

This work was funded in part by a grant from the National Institute of Allergy and Infectious Diseases (5T32AI007532-24 to OAP). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.