The usefulness of nanopore sequencing in whole-genome sequencing-based genotyping of Listeria monocytogenes and Salmonella enterica serovar Enteritidis

Microbiol Spectr. 2024 Jul 2;12(7):e0050924. doi: 10.1128/spectrum.00509-24. Epub 2024 May 29.

Abstract

Bacterial genotyping through whole-genome sequencing plays a crucial role in disease surveillance and outbreak investigations in public health laboratories. This study assessed the effectiveness of Oxford Nanopore Technologies (ONT) sequencing in the genotyping of Listeria monocytogenes and Salmonella enterica serovar Enteritidis. Our results indicated that ONT sequences, generated with the R10.4.1 flow cell and basecalled using the Dorado 0.5.0 Super Accurate 4.3 model, exhibited comparable accuracy to Illumina sequences, effectively discriminating among bacterial strains from outbreaks. These findings suggest that ONT sequencing has the potential to be a promising tool for rapid whole-genome sequencing of bacterial pathogens in public health laboratories for epidemiological investigations.

Importance: This study unveils that Oxford Nanopore Technologies sequencing, by itself, holds the potential to serve as a whole-genome sequencing-based genotyping tool in public health laboratories, enabling routine subtyping of bacterial isolates for disease surveillance and outbreak investigations.

Keywords: Listeria monocytogenes; ONT sequencing; Salmonella enterica serovar Enteritidis; cgMLST; genotyping; wgSNP; whole-genome sequencing.

MeSH terms

  • Disease Outbreaks
  • Genome, Bacterial* / genetics
  • Genotype
  • Genotyping Techniques / methods
  • Humans
  • Listeria monocytogenes* / classification
  • Listeria monocytogenes* / genetics
  • Listeria monocytogenes* / isolation & purification
  • Listeriosis / microbiology
  • Nanopore Sequencing* / methods
  • Salmonella Infections / microbiology
  • Salmonella enteritidis* / classification
  • Salmonella enteritidis* / genetics
  • Salmonella enteritidis* / isolation & purification
  • Whole Genome Sequencing* / methods