Abstract

Norovirus is estimated to cause 677 million annual cases of gastroenteritis worldwide, resulting in 210,000 deaths. As viral gastroenteritis is generally self-limiting, clinical samples for epidemiological studies only partially represent circulating noroviruses in the population and is biased towards severe symptomatic cases. As infected individuals from both symptomatic and asymptomatic cases shed viruses into the sewerage system at a high concentration, waste water samples are useful for the molecular epidemiological analysis of norovirus genotypes at a population level. Using Illumina MiSeq and Sanger sequencing, we surveyed circulating norovirus within Australia and New Zealand, from July 2014 to December 2016. Importantly, norovirus genomic diversity during 2016 was compared between clinical and waste water samples to identify potential pandemic variants, novel recombinant viruses and the timing of their emergence. Although the GII.4 Sydney 2012 variant was prominent in 2014 and 2015, its prevalence significantly decreased in both clinical and waste water samples over 2016. This was concomitant with the emergence of multiple norovirus strains, including twoGII.4 Sydney 2012 recombinant viruses, GII.P4 New Orleans 2009/GII.4 Sydney 2012 and GII.P16/GII.4 Sydney 2012, along with three other emerging strains GII.17, GII.P12/GII.3 and GII.P16/GII.2. This is unusual, as a single GII.4 pandemic variant is generally responsible for 65–80% of all human norovirus infections at any one time and predominates until it is replaced by a new pandemic variant. In sumary, this study demonstrates the combined use of clinical and wastewater samples provides a more complete picture of norovirus circulating within the population.

Highlights

  • Next-generation sequencing (NGS) is a new and rapidly evolving technology that facilitates the simultaneous sequencing of large amounts of genetic material

  • The prevalence of circulating norovirus GI and GII genotypes in Australia and New Zealand (NZ) were determined in this study, which included a total of 782 norovirus-positive specimens

  • To compare the identified norovirus sequences, a subset of 89 Australian (GI = 11 and GII = 78) and 104 NZ (GI = 33 and GII = 71) representative sequences were selected for phylogenetic tree construction (Figs. 1 and 2)

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Summary

Introduction

Next-generation sequencing (NGS) is a new and rapidly evolving technology that facilitates the simultaneous sequencing of large amounts of genetic material. Molecular epidemiological studies of norovirus are usually performed with clinical samples collected from patients presenting at medical facilities. This is not representative of the whole norovirus population and is biased towards severe symptomatic cases, offering only a narrow picture of the complete viral diversity. Institutional outbreaks of viral gastroenteritis are difficult to control and have significant global economic burden to public healthcare systems (USD $4.2 billion) and communities (USD $60.3 billion) each year[6]

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