Abstract

Whole-genome sequencing (WGS) has emerged as a powerful tool for comparing bacterial isolates in outbreak detection and investigation. Here we demonstrate that WGS performed prospectively for national epidemiologic surveillance of Listeria monocytogenes has the capacity to be superior to our current approaches using pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), multilocus variable-number tandem-repeat analysis (MLVA), binary typing, and serotyping. Initially 423 L. monocytogenes isolates underwent WGS, and comparisons uncovered a diverse genetic population structure derived from three distinct lineages. MLST, binary typing, and serotyping results inferred in silico from the WGS data were highly concordant (>99%) with laboratory typing performed in parallel. However, WGS was able to identify distinct nested clusters within groups of isolates that were otherwise indistinguishable using our current typing methods. Routine WGS was then used for prospective epidemiologic surveillance on a further 97 L. monocytogenes isolates over a 12-month period, which provided a greater level of discrimination than that of conventional typing for inferring linkage to point source outbreaks. A risk-based alert system based on WGS similarity was used to inform epidemiologists required to act on the data. Our experience shows that WGS can be adopted for prospective L. monocytogenes surveillance and investigated for other pathogens relevant to public health.

Highlights

  • Listeria monocytogenes is a predominantly food-borne pathogen capable of causing a range of clinical illnesses, including invasive disease such as bacteremia and meningoencephalitis in humans, and is commonly monitored by public health facilities for the emergence of outbreaks [1, 2]

  • We evaluated the use of routine prospective Whole-genome sequencing (WGS) compared with the use of conventional typing methods, including pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), multilocus variable-number tandem-repeat analysis (MLVA), binary typing, and PCR serotyping, for the national epidemiologic surveillance of L. monocytogenes

  • Based on a phylogeny inferred from an alignment of 158,707 core genome single nucleotide polymorphisms (SNPs), WGS revealed an Australian L. monocytogenes population structure derived from three distinct evolutionary lineages (Fig. 1)

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Summary

Introduction

Listeria monocytogenes is a predominantly food-borne pathogen capable of causing a range of clinical illnesses, including invasive disease such as bacteremia and meningoencephalitis in humans, and is commonly monitored by public health facilities for the emergence of outbreaks [1, 2]. Whole-genome sequencing (WGS) has emerged as a powerful technology for the comparison of isolates in outbreak analysis. The Microbiological Diagnostic Unit Public Health Laboratory is the Australian Listeria reference laboratory and routinely performs molecular typing of human and nonhuman isolates of L. monocytogenes referred from local and interstate laboratories. We evaluated the use of routine prospective WGS compared with the use of conventional typing methods, including pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), multilocus variable-number tandem-repeat analysis (MLVA), binary typing, and PCR serotyping, for the national epidemiologic surveillance of L. monocytogenes

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