Global Trends in Norovirus Genotype Distribution among Children with Acute Gastroenteritis.

Noroviruses are a leading cause of acute gastroenteritis (AGE) among adults and children worldwide. NoroSurv is a global network for norovirus strain surveillance among children <5 years of age with AGE. Participants in 16 countries across 6 continents used standardized protocols for dual typing (genotype and polymerase type) and uploaded 1,325 dual-typed sequences to the NoroSurv web portal during 2016-2020. More than 50% of submitted sequences were GII.4 Sydney[P16] or GII.4 Sydney[P31] strains. Other common strains included GII.2[P16], GII.3[P12], GII.6[P7], and GI.3[P3] viruses. In total, 22 genotypes and 36 dual types, including GII.3 and GII.20 viruses with rarely reported polymerase types, were detected, reflecting high strain diversity. Surveillance data captured in NoroSurv enables the monitoring of trends in norovirus strains associated childhood AGE throughout the world on a near real-time basis.

GII.4 norovirus recombinant causes gastroenteritis epidemic in Eastern Australia, 2017.

In Victoria, Australia, 160 gastroenteritis outbreaks were norovirus positive for the period January-September 2017. A distinctive peak in norovirus outbreaks was seen May-August, with 118 positive outbreaks occurring in the peak period. The peak was primarily due to the emergence of a GII.P4_NewOrleans_2009/GII.4_Sydney_2012 recombinant that had genetically changed sufficiently to escape herd immunity. This recombinant was also identified elsewhere in Australia, with highly similar sequences identified in Queensland during the same time period. The recombinant GII.P4_NewOrleans_2009/GII.4_Sydney_2012 has not been reported to cause norovirus epidemics outside Australia, suggesting regional factors play a role in determining norovirus genotype incidence.

The molecular epidemiology of norovirus outbreaks in Victoria, 2014 to 2015
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Noroviruses are a leading cause of outbreaks of gastroenteritis. This study examined the incidence and molecular characteristics of norovirus outbreaks in healthcare and non-healthcare settings in Victoria, Australia, over 2 years (2014-2015). Norovirus was detected in 65.7% and 60.4% of gastroenteritis outbreaks investigated for the years 2014 and 2015 respectively. There was a significant decline in the number of norovirus outbreaks in the period 2014 to 2015 although in both years norovirus outbreaks peaked in the latter part of the year. Norovirus Open Reading Frame (ORF) 2 (capsid) genotypes identified included GI.2, GI.3, GI.4, GI.5, GI.6, GI.9, GII.2, GII.3, GII.4, GII.6, GII.7, GII.8, GII.13 and GII.17. GII.4 was the most common genotype detected. In addition, the following ORF 1/ORF 2 recombinant forms were confirmed: GII.P4_NewOrleans_2009/GII.4_Sydney_2012, GII.P12/GII.3, GII.Pb (GII.21)/GII.3, GII.Pe/GII.2 and GII.Pe/GII.4_Sydney_2012. A significant decline was noted in the chief norovirus strain GII.Pe/GII.4_Sydney_2012 between 2014 and 2015 but there was a re-emergence of a GII.P4_ NewOrleans _2009 norovirus strain. Outbreaks involving the GII.P17/GII.17 genotype were also detected for the first time in Victoria. GI genotypes circulating in Victoria for the 2 years 2014 and 2015 underwent a dramatic change between the 2 years of the survey. Many genotypes could occur in both healthcare and non-healthcare settings although GI.3, GII.6, and GII.4 were significantly more common in healthcare settings. The study emphasises the complex way in which norovirus circulates throughout the community.

Emergence of GII.Pg norovirus in gastroenteritis outbreaks in Victoria, Australia.

The ORF 1 GII.Pg genotype represents an obligatory recombinant comprising the ORF 1 GII.Pg genotype and a number of ORF 2 genotypes. The emergence, incidence, and molecular features of GII.Pg norovirus have never been considered in detail and are the subject of the current study. Over the period 2002-2013, GII.Pg norovirus was detected in 16 outbreaks in Victoria, Australia. It was first identified in 2009 and thereafter was detected at low level in each year of the study. GII.Pg norovirus outbreaks occurred in both healthcare and non-healthcare settings and could be found in individuals with a broad range of ages. The seasonality of GII.Pg norovirus outbreaks was significantly different from that of all other (non-GII.Pg) norovirus outbreaks. For the 15 GII.Pg norovirus outbreaks where ORF 2 sequencing data were available, two ORF 2 recombinant genotypes were found: GII.1 in 5 (33%) outbreaks and GII.12 in 10 (67%) outbreaks. The ORF 1 phylogenetic tree shows that the GII.Pg ORF 1 genotype fell into two distinct groups. The ORF 2 phylogenetic tree indicates that the GII.1 and GII.12 clusters each corresponded to one of the groups in the ORF 1 tree. This indicates the two recombinant forms were evolving in parallel and not one from the other. Analysis of age data indicates the GII.1 and GII.12 recombinant forms circulated in different ways in the community. J. Med. Virol. 88:1521-1528, 2016. © 2016 Wiley Periodicals, Inc.

The Comparative Molecular Epidemiology of GII.P7_GII.6 and GII.P7_GII.7 Norovirus Outbreaks in Victoria, Australia, 2012-2014.

The comparative molecular epidemiology of the related GII.P7_GII.6 and GII.P7_GII.7 noroviruses has not been examined in detail. ORF 1, ORF 2 and ORF 1/ORF 2 RT-PCR as well as sequencing and phylogeny analysis were carried out on faecal specimens from 873 gastroenteritis outbreaks in Victoria, Australia (2012-2014). There were 575 (66%) detected as positive for norovirus by means of ORF 1 RT-PCR and/or ORF 2 RT-PCR. Of these, 24 (4.2%) were GII.6 (ORF 2) outbreaks, 7 (1.2%) were GII.7 (ORF 2) outbreaks, and 1 outbreak (0.2%) involved both GII.6 (ORF 2) and GII.7 (ORF 2) noroviruses. The median age of patients identified with GII.6 (ORF 2) (84 years) was significantly different from that of patients identified with GII.7 (ORF 2) (39 years). ORF 2 GII.6 and ORF 2 GII.7 sequences were always associated with a GII.P7 ORF 1 sequence, and GII.P7 sequences fell into two clusters, with one corresponding to the GII.6 ORF 2 genotype and the other to the GII.7 ORF 2 genotype, thereby indicating that the ORF 1 has been evolving separately for the two viruses. Thus, two closely related noroviruses can have a markedly different incidence and epidemiology.

A norovirus intervariant GII.4 recombinant in Victoria, Australia, June 2016: the next epidemic variant?

A norovirus recombinant GII.P4_NewOrleans_2009/GII.4_Sydney_2012 was first detected in Victoria, Australia, in August 2015 at low frequency, and then re-emerged in June 2016, having undergone genetic changes. Analysis of 14 years' surveillance data from Victoria suggests a typical delay of two to seven months between first detection of a new variant and occurrence of a subsequent epidemic linked to that variant. We consider that the current recombinant strain has the potential to become a pandemic variant.

Norovirus genotype diversity in community-based sporadic gastroenteritis incidents: a five-year study.

Although norovirus is a known cause of sporadic gastroenteritis, the incidence and genotypes of norovirus associated with sporadic community-based gastroenteritis are poorly understood. The current study examined this issue by using material from alleged food poisoning incidents in the state of Victoria, Australia, for the period 2008-2012. Norovirus was identified, by either ORF (open reading frame) 1 or ORF 2 RT-PCR methodology, in 159 of 379 (42.0%) sporadic gastroenteritis incidents, thereby showing that norovirus was an important cause of sporadic gastroenteritis. The number of sporadic norovirus incidents did not vary significantly from year to year, indicating that the pool of circulating norovirus remained constant. Norovirus ORF 1 genotypes identified included GI.1, GI.2, GI.3, GI.4, GI.b, GI.d, GII.2, GII.4 (including variants 2006a, 2006b, 2007, and 2009), GII.16, GII.22, GII.b, GII.e, and GII.g. Norovirus ORF 2 genotypes identified included GI.1, GI.2, GI.3, GI.4, GI.6, GII.2, GII.3, GII.4 (variants 2006b, 2009, 2009-like, 2012, and "unknown"), GII.6, GII.7, GII.9, GII.12, and GII.13. Five ORF 1/ORF 2 norovirus recombinant forms were confirmed: GII.b/GII.3, GII.e/GII.4 (2012), GII.e/GII.4 (unknown), GII.g/GII.12 and GII.16/GII.2. Although the incidence of ORF 2 GI.3 was significantly higher in children than in adults, this was not the case for other major ORF 2 genotypes (GII.2, GII.4, and GII.6) which occurred equally in all age groups. The findings demonstrate the importance and diverse nature of norovirus in sporadic community-based gastroenteritis incidents and indicate that the development of successful vaccine strategies may be difficult.

Norovirus genotype diversity associated with gastroenteritis outbreaks in Victoria in 2013.

The noroviruses are now considered a leading cause of outbreaks of non-bacterial gastroenteritis worldwide. Vaccine strategies against norovirus are currently under consideration but depend on a detailed knowledge of the capsid genotypes. This study examined the incidence of norovirus outbreaks in Victoria over 1 year (2013) and documented the genotypes occurring in the different outbreak settings (healthcare and non-healthcare) and age groups. It was found that 63.1% of gastroenteritis outbreaks were associated with norovirus, thereby showing norovirus to be the major viral cause of illness in gastroenteritis outbreaks. Sixteen capsid genotypes were identified and included GI.2, GI.3, GI.4, GI.6, GI.7, GI.8, GI.9, GII.1, GII.2, GII.3, GII.4, GII.5, GII.6, GII.7, GII.13 and an as yet unclassified GII genotype. All genotypes found in the study, with the exception of GI.9, were detected in the elderly, indicating prior exposure to a norovirus genotype did not appear to confer long term immunity in many cases. The incidence of genotypes GII.1, GII.4 and GII.7 was linked with setting and age. As setting and age were correlated it was not possible to determine which variable was critical with the exception of GII.7, which appeared to be linked to age. The findings indicate that norovirus vaccine strategies should encompass a broad range of genotypes and, as setting or age may be important in determining genotype incidence, this should be taken into account as well.

Changes in norovirus incidence in Victoria, Australia, 2022: are we back to normal yet after COVID?

Abstract: There were 108 norovirus-positive outbreaks in 2022, with 45 (41.7%) occurring during the first quarter (Q1), January-March. Aged care facilities accounted for 44.4% of norovirus-positive outbreaks; 43.5% were in childcare settings. Overall, the GII.P31/GII.4 genotype was the most common, involved in 39.4% of outbreaks; however, there were shifts in the most common genotype across the year. In Q1, the GII.P31/GII.4 genotype accounted for 73.3% of typed outbreaks, but by Q3 (July-September) the GII.P7/GII.6 was the most prominent genotype at 45.0%. In Q4 (October-December), the dominant genotype had changed again to GII.P16/GII.4 (52.6%). While the incidence of norovirus outbreaks in 2022 was average regarding overall prevalence and genotype diversity, there are still ongoing effects from the coronavirus disease 2019 (COVID-19) pandemic in relation to seasonality, outbreak demographics and specimen referral.

Australian National Enterovirus Reference Laboratory annual report, 2022.

Abstract: Australia monitors its polio-free status by conducting surveillance for cases of acute flaccid paralysis (AFP) in children less than 15 years of age, as recommended by the World Health Organization (WHO). Cases of AFP in children are notified to the Australian Paediatric Surveillance Unit or the Paediatric Active Enhanced Disease Surveillance System, and faecal specimens are referred for virological investigation to the National Enterovirus Reference Laboratory. In 2022, no cases of poliomyelitis were reported from clinical surveillance and Australia reported 1.69 non-polio AFP cases per 100,000 children, thereby meeting the WHO's performance criterion for a sensitive surveillance system. The non-polio enteroviruses coxsackievirus A2, coxsackievirus A6, coxsackievirus A10, echovirus 18, enterovirus A71 and enterovirus C96 were identified from clinical specimens collected from AFP cases. Australia also performs enterovirus and environmental surveillance to complement the clinical system focussed on children. In 2022, thirty cases of wild poliovirus were reported from three countries (Afghanistan, Mozambique and Pakistan); 24 countries also reported cases of poliomyelitis due to circulating vaccine-derived poliovirus.

Molecular and epidemiological features of gastroenteritis outbreaks involving genogroup I norovirus in Victoria, Australia, 2002-2010.

GI noroviruses are relatively rare and systematic studies of the molecular epidemiology of GI norovirus outbreaks are lacking. The current study examined the molecular virology of GI norovirus outbreaks in Victoria, Australia (2002-2010). Of 1,617 norovirus outbreaks identified, 69 (4.3%) were associated with GI norovirus alone, 1,540 (95.2%) with GII norovirus alone and 8 (0.5%) with GI + GII. Some differences between GI and GII outbreak epidemiology were found. GI outbreaks peaked in the 2-month period November/December whereas GII outbreaks peaked in the 2-month period September/October and GI norovirus outbreaks were significantly more common in non-healthcare settings (37.7%) than GII outbreaks (9.5%). ORF 1/ORF 2 genotypes found in the 69 outbreaks involving GI norovirus alone were: GI.2/GI.2, 7 outbreaks; GI.2/GI.6, 18 outbreaks; GI.3b/GI.3, 14 outbreaks; GI.4/GI.4, 21 outbreaks; GI.8/GI.8, one outbreak; GI.d/GI.3, four outbreaks; and GI.e/GI.13, one outbreak. The current study appears to be the first to have identified the recombinant form, GI.2/GI.6. Whereas GI.2/GI.6 and GI.3b/GI.3 outbreaks occurred with equal frequency in both healthcare and non-healthcare settings, GI.4/GI.4 occurred predominantly in healthcare settings. GI ORF 1/ORF 2 genotypes found in the eight outbreaks involving GI + GII norovirus were GI.2/GI.6, GI.3b/GI.3, and GI.4/GI.4, indicating GI genotypes in GI + GII outbreaks were similar to those found in outbreaks involving GI alone. Apparent differences in the evolution of different GI genotypes were noted. GI.2/GI.2, GI.2/GI.6, and GI.4/GI.4 strains tended to undergo periodic shifts in nucleotide sequence whereas various GI.3b/GI.3 strains tended to circulate simultaneously.

Changes in norovirus incidence in Victoria, Australia, during the COVID-19 pandemic, 2020-2021.

There were 142 norovirus positive outbreaks in Victoria for the 2020-2021 calendar years; however, almost half of these (48.6%) occurred in Q1 (January-March) of 2021. For the two-year period, 69.0% of all norovirus positive outbreaks were in childcare settings, and the predominant genotype was GII.P16/GII.2 (64.9%) followed by GII.P31/GII.4_2012 (20.9%). Norovirus incidence was particularly low in 2020 (n = 26) and close to average in 2021 (n = 116), but genotype diversity was low in both years. With the thought that 2022 will approach a more normal aspect to socialising and travel, norovirus incidence in 2022 may be predicted to increase above typical levels.

Australian National Enterovirus Reference Laboratory annual report, 2021.

Australia monitors its polio-free status by conducting surveillance for cases of acute flaccid paralysis (AFP) in children less than 15 years of age, as recommended by the World Health Organization (WHO). Cases of AFP in children are notified to the Australian Paediatric Surveillance Unit or the Paediatric Active Enhanced Disease Surveillance System, and faecal specimens are referred for virological investigation to the National Enterovirus Reference Laboratory. In 2021, no cases of poliomyelitis were reported from clinical surveillance and Australia reported 1.31 non-polio AFP cases per 100,000 children, thereby meeting the WHO's performance criterion for a sensitive surveillance system. The non-polio enteroviruses coxsackievirus A4, coxsackievirus A10, coxsackievirus A13 and enterovirus A71 were identified from clinical specimens collected from AFP cases. Australia also performs enterovirus and environmental surveillance to complement the clinical system focussed on children. In 2021, there were five cases of wild poliovirus reported from the two remaining endemic countries: Afghanistan and Pakistan. Including Afghanistan and Pakistan, 22 countries also reported cases of AFP due to circulating vaccine-derived poliovirus.

Australian National Enterovirus Reference Laboratory annual report, 2020.

ABSTRACT: Australia monitors its polio-free status by conducting surveillance for cases of acute flaccid paralysis (AFP) in children less than 15 years of age, as recommended by the World Health Organization (WHO). Cases of AFP in children are notified to the Australian Paediatric Surveillance Unit or the Paediatric Active Enhanced Disease Surveillance System and faecal specimens are referred for virological investigation to the National Enterovirus Reference Laboratory. In 2020, no cases of poliomyelitis were reported from clinical surveillance; Australia reported 1.09 non-polio AFP cases per 100,000 children, thereby meeting the WHO's performance criterion for a sensitive surveillance system. The non-polio enteroviruses coxsackievirus A10 and coxsackievirus A16 were identified from clinical specimens collected from AFP cases. Australia also performs enterovirus surveillance and environmental surveillance to complement the clinical system focussed on children. In 2020, there were 140 cases of wild poliovirus reported from the two remaining endemic countries: Afghanistan and Pakistan. Another 28 countries reported cases of circulating vaccine-derived poliovirus.

Decreased incidence of enterovirus and norovirus infections during the COVID-19 pandemic, Victoria, Australia, 2020.

ABSTRACT: Significant reductions in the incidence of enteroviruses and noroviruses, both transmitted primarily by the faecal-oral route, were noted in 2020 compared to the previous decade, in Victoria, Australia. The enterovirus specimen positivity rate was reduced by 84.2% in 2020, while the norovirus outbreak positivity rate declined by 49.0%. The most likely explanation for these reductions is the concurrence of social restrictions, physical distancing, personal hygiene awareness and international and domestic border closures in response to the COVID-19 pandemic.

Molecular changes associated with altered patterns of norovirus outbreak epidemics in Victoria, Australia, in 2006 to 2007.

Noroviruses (NoVs) are now considered the most common cause of outbreaks of nonbacterial gastroenteritis, but the factors which control the incidence of NoVs are poorly understood. In 2006, the pattern of NoV outbreak epidemics in Victoria, Australia, changed compared to the pattern for 2002 to 2005 and 2007. This study examined molecular correlates of the changed NoV periodicity. For the period of 2002 to 2007, 8,507 fecal specimens from 1,495 gastroenteritis outbreaks were tested for NoV by reverse transcription-PCR, and 1,018 NoV outbreaks were identified. Nucleotide sequence analysis was used to define genotypes and GII.4 variants. For 2002 to 2007, GII.4 was the predominant genotype. For the period of 2002 to 2005 and 2007, a single NoV outbreak epidemic occurred in warmer months of each year, but in 2006 two epidemics occurred in 1 year, one in colder months and one in warmer months of the year. For 2002 to 2007, four major GII.4 variants, "2002 Oxford/Farmington Hills," "2004 Hunter," "2006a," and "2006b," were identified. Each NoV outbreak epidemic was linked principally to one of these four variants, and there was a time link, a delay of 2 to 6 months, between the first detection of a GII.4 variant and the first outbreak epidemic in which it was the principal variant. The unusual 2006 pattern of outbreak epidemics can then be correlated with the appearance of two GII.4 variants within a short space of time, resulting in two outbreak epidemics in a short space of time, i.e., in the 1 year. This study provides a potentially greater ability to predict the characteristics of NoV epidemics.

Molecular and epidemiological characteristics of norovirus associated with community-based sporadic gastroenteritis incidents and norovirus outbreaks in Victoria, Australia, 2002-2007.

OBJECTIVES: The molecular and epidemiological features of community-based norovirus-associated sporadic gastroenteritis incidents (NASGIs) are poorly understood. This study examined these features and compared the findings with studies of community-based and institutional norovirus-associated gastroenteritis outbreaks (NAGOs). METHODS: Fecal specimens from NASGIs and NAGOs that occurred in Victoria, Australia (2002-2007) were tested for norovirus by reverse transcription-polymerase chain reaction methodology. Norovirus genotype was determined by nucleotide sequence analysis. RESULTS: 106 community-based NASGIs, 116 community-based NAGOs and 902 institutional NAGOs were identified. The mean age and gender ratio of individuals associated with community-based NASGIs and community-based NAGOs were similar but differed from that found for institutional NAGOs. Although GII.4 was the predominant genotype associated with all three incident types, the mix of genotypes was similar for community-based NASGIs and community-based NAGOs but that for institutional NAGOs was different. All three incident types had a similar seasonal periodicity due to the pronounced seasonal periodicity of GII.4 incidents. CONCLUSIONS: The molecular and epidemiological features of noroviruses associated with community-based NASGIs and community-based NAGOs are similar but are different from those found for institutional NAGOs.

The molecular epidemiology of norovirus outbreaks in Victoria, 2016.

Noroviruses are a leading cause of outbreaks of gastroenteritis. This study examined the incidence and molecular characteristics of norovirus outbreaks in Victoria, Australia in 2016. Norovirus was detected in 52.4% of gastroenteritis outbreaks surveyed and the year was notable in that there was no significant temporal peak in norovirus outbreaks. Norovirus Open Reading Frame (ORF)2 (capsid) genotypes were successfully identified in 84 of 110 norovirus outbreaks and included GI.3, GI.6, GI.9, GII.2, GII.3, GII.4, GII.6, and GII.17. Norovirus GII.4 was the most common ORF2 genotype detected (55.9%). Other relatively common ORF2 genotypes included GII.2 (19.0%), GII.17 (9.5%), GI.3 (7.1%) and GII.3 (4.8%). The GII.P4_NewOrleans_2009/GII.4_Sydney_2012 recombinant emerged as an important new GII.4 form. The study also confirmed the identity of three ORF1/ORF2 recombinant forms as follows: GII.P16/GII.2, GII.P16/GII.3 and GII.P16/GII.4. Statistical analysis indicated GII.4 (ORF2) was much more common in healthcare settings than in non-healthcare settings. The study indicates 2016 was a transition year in Victoria, Australia, in that the previous norovirus epidemic strain had diminished to the point where it was no longer dominant but as yet no replacement epidemic strain had become obvious.

Evolutionary changes in the capsid P2 region of Australian strains of the norovirus GII.Pe_GII.4.

PURPOSE: The protruding (P) 2 region of the norovirus capsid is thought to include hypervariable sites involved in receptor binding. This study examines the changes that occurred in the P2 region of GII.Pe_GII.4 norovirus in the course of its evolution from a precursor phase (2008-2009), to an intermediate phase (2010) and finally to an epidemic phase (2012-2015). METHODOLOGY: Twenty-two P2 region amino acid (aa) sequences (166 aa long) from all phases of the evolution of the virus were compared and the changes analysed.Results/key findings. Twenty sites in the P2 region underwent aa change and of these, 10 corresponded to previously proposed hypervariable sites and 10 to novel hypervariable sites. It was notable that aa changes at two sites, X and Y, only emerged as the epidemic phase progressed. 3D computer modelling of the P2 region indicated that neither X nor Y were in the uppermost 'crown', but further down in the 'neck' portion. The location of X and Y and the nature of aa change at Y suggest these sites were important in enhancing the structural integrity of the capsid, which in turn may have facilitated the longer term viability of the virus. CONCLUSION: The current study helps establish the validity of previously proposed hypervariable sites in the P2 region as well as indicating new ones. It also provides quantitative and qualitative data on how these sites changed over the evolutionary history of a particular norovirus strain.