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The emergence of the novel GII.17 Kawasaki 2014 norovirus variant raising the interest of the public, has replaced GII.4 as the predominant cause of noroviruses outbreaks in East Asia during 2014–2015. Antigenic variation of the capsid protein is considered as one of the key mechanisms of norovirus evolution. In this study, we screened a panel of GII.17 mutants. First, we produced norovirus P proteins using cell-free protein synthesis (CFPS) system, comparing the results to pure proteins expressed in a cell-based system. Next, we determined the binding capability of specific monoclonal antibody (mAb) 2D11 using a unique set of wild-type GII.17 strains. Results of the EIA involving a panel of mutant cell-free proteins indicated that Q298 was the key residue within loop 1. These data highlighted the essential residues in the linear antibody binding characteristics of novel GII.17. Furthermore, it supported the CFPS as a promising tool for rapidly screening mutants via the scalable expression of norovirus P proteins. [ABSTRACT FROM AUTHOR]

Human norovirus is regarded as the leading cause of epidemic acute gastroenteritis with GII.4 being the predominant genotype during the past decades. In the winter of 2014/2015, the GII.17 Kawasaki 2014 emerged as the predominant genotype, surpassing GII.4 in several East Asian countries. Hence, the influence of host immunity response on the continuous evolution of different GII.17 variants needs to be studied in depth. Here, we relate the inferences of evolutionary mechanisms of different GII.17 variants with the investigation of cross-reactivity and cross-protection of their respective antisera using the expression of norovirus P particles in Escherichia coli. The cross-reactivity assay showed that the antisera of previous strains (GII.17 A and GII.17 B) reacted with recent variants (GII.17 C and GII.17 D) at high OD values from 0.8 to 1.16, while recent variant antisera cross-reacting with previous strains were weak with OD values between 0.26 and 0.56. The cross-protection assay indicated that the antisera of previous strains had no inhibitory effect on recent variants. Finally, mutations at amino acids 353–363, 373–384, 394–404, and 444–454 had the greatest impact on cross-reactivity. These data indicate that the recent pandemic variants GII.17 C and GII.17 D avoided the herd immunity effect of previous GII.17 A and GII.17 B strains through antigenic variation. [ABSTRACT FROM AUTHOR]

Abstract Noroviruses are the primary cause of non-bacterial acute gastroenteritis worldwide, and GII.8 belongs to a non-epidemic genotype with a limited understanding currently. In this study, we assembled the first GII.8 norovirus genome from China and clarified the temporal evolutionary process of this non-epidemic variant. Using the "4+1+1" application strategy with newly designed primer sets, the genome of one GII.8 strain GZ2017-L601 from China was firstly sequenced that comprised 7476 nucleotides. The homology of the new genome and the previous only GII.8 genome reached 93.8% identity at the nucleotide level, but only 10, 6, 7 amino acid mutations occurred in three ORFs. When compared the new strain with other GII reference strains, p22 and P2 were calculated as the variable encoding regions, and NTPase, VPg, 3CL, RdRp and S were shown as the conserved ones. We then reconstructed the evolutionary process of the GII.8 genotype using other available sequences in GenBank. Based on the partial N/C region, all GII.8 strains could be subdivided chronologically into four clusters, which spans 1967–1994, 1997–2005, 2003–2009, and 2007–2017, respectively. Moreover, differences of capsid P proteins between GII.8 strains and the epidemic GII.4 strain VA387 were also compared. There existed 147/310 distinct amino acid sites in the alignment, including two insertion and three deletion mutations. Distribution of antigen epitopes of two GII.8 variants was comparable, but the numbers of antigenic sites of GII.8 strains were less than that of VA387. In summary, the first GII.8 genome from China was assembled and extensively characterized, and a time-order evolutionary process of this genotype was identified with a static pattern of antigenic variations. Highlights • The first GII.8 norovirus genome sequence from China was obtained and analyzed. • GII.8 could be chronologically divided into four clusters based on the N/C region. • The numbers of antigenic sites of GII.8 strains were less than that of GII.4 VA387. • No obvious changes in epitope distribution were found between different GII.8 strains. [ABSTRACT FROM AUTHOR]

Background: Human norovirus is regarded as the leading cause of nonbacterial acute diarrhea in developing and developed countries. Among all genotypes, GII.4 has been the predominant genotype, but in East Asia, it was replaced by the GII.17 in 2014/2015. However, after the prevalence of new GII.17 variant in South China, a sharply increase in the number of norovirus infections associated with sporadic acute diarrhea was detected. In this study, we would investigate the prevalence and genetic diversity of noroviruses in the sporadic acute gastroenteritis cases in the post-GII.17 period in South China.Methods: Norovirus was screened from 217 patients with sporadic acute gastroenteritis from August 2015 to October 2017 by reverse transcription-polymerase chain reaction. Then, two regions including the partial RNA polymerase and the capsid gene of positive samples were amplified and sequenced. Phylogenetic analyses were performed to determine norovirus genotypes. Complete VP1 sequences of GII.4 strains detected in this study were also amplified and subjected into evolutionary tracing analyses.Results: A total of 43 (19.82%) norovirus samples were confirmed from 217 stool specimens, and it was found that GII.4 resurged as the new predominant variant, accounting for 76.74% (33/43) of positive samples. Only one local strain GZ2015-L550 was clustered with the contemporary GII.P16/GII.4-2012 recombinant variant, and other 32 local strains belonged to the clade with the GII.Pe/GII.4-2012 variant. Other genotypes including GII.17 (n = 4), GII.3 (n = 4), GII.8 (n = 1) and GI. 6 (n = 1) were also detected. Furthermore, all GII.4 strains were phylogenetic analyzed based on their capsid P2 subdomains. Combined with other reported 754 strains, the GII.4-2012 variant could be divided into two clades. Most GII.4 strains collected in 2016 and 2017 in this study (7/8) formed a new cluster A in Clade II with additional 103 contemporaneous strains. In addition, evolutionary tracing of the capsid P2 subdomain of this variant was also analyzed, and one specific amino acid substitutions (N373) was identified for Cluster A.Conclusion: In summary, this study confirmed a norovirus infection peak in the post-GII.17 period in South China, which was caused by the resurgence of the GII.4 variant. [ABSTRACT FROM AUTHOR]

Human noroviruses (HuNoVs) have been found as the leading cause of acute gastroenteritis outbreaks in all age groups and are significantly correlated with the consumption of shellfish. In this study, the contamination of HuNoVs in shellfish was estimated through a systematic review and meta-analysis. Studies on the contamination of HuNoVs in shellfish were searched from PubMed, Web of Science, Embase, and Cochrane Library from January 2000 to August 2021. A total of 75 studies were included, and the pooled HuNoVs prevalence in shellfish was 29% (95% CI: 23–35) worldwide. As revealed by the results of the subgroup meta-analysis, the prevalence of dominant genogroup was variable, and 4% (95% CI: 3–6), 13% (95% CI: 10–17), with 7% (95% CI: 4–11) of the samples, respectively, contaminated by GI alone, GII alone, and GI&GII. The HuNoVs prevalence of shellfish in Europe, America, and Asia was 33% (95% CI: 24–43), 24% (95% CI: 7–47), and 27% (95% CI: 18–35), respectively, while only 10% (95% CI: 5–17) in Africa. Furthermore, the prevalence of HuNoVs in shellfish was the highest in spring (35%, 95% CI: 23–49) and winter (35%, 95% CI: 22–50), and the lowest in summer (11%, 95% CI: 5–18). Oysters, clams, and mussels had comparable HuNoVs prevalence of 28% (95% CI: 20–37), 27% (95% CI: 16–39) and 24% (95% CI: 17–32), respectively. The prevalence of HuNoVs in shellfish from harvest areas and markets was 30% (95% CI: 23–38) and 30% (95% CI: 19–41), respectively. The results of this study suggest a substantial burden of HuNoVs in shellfish worldwide, with GII.4 (92.86%) and GII.2 (46.43%) as the predominant genotypes. This study provides information regarding the contamination of HuNoVs in shellfish worldwide, which will contribute to the development of appropriate control measures to prevent shellfish-related HuNoVs gastroenteritis. • The pooled prevalence of human noroviruses in shellfish was found as 29% (95% CI: 23–35) worldwide. • GII.4 (92.86%, 26/28) was found as the most common genotype in shellfish. • Significant differences were found in the seasonal and geographical prevalence of human noroviruses in shellfish. [ABSTRACT FROM AUTHOR]

Noroviruses (NoVs) are the primary cause of non-bacterial acute gastroenteritis worldwide. Most NoV infections are caused by GII.4, but GII.6 is also an important genotype with a long-term persistence in human populations. In this study, the complete genome sequence of a NoV strain GZ2010-L96 isolated in China was identified and analyzed phylogenetically. The viral genome comprised 7550 nucleotides, and its phylogenetic analysis revealed that the strain belonged to GII.6 genotype. All reported GII.6 NoV capsid protein sequences were also collected for comparative analysis, and GZ2010-L96 was clustered into GII.6-b with other 8 strains. Meanwhile, it was found that 53 spots on viral capsid showed subcluster specificity according to multiple alignments. Moreover, homologous modeling of GZ2010-L96 based on comparison with GII.4 VA387 strain showed a different antigen distribution pattern. In summary, the genome of the GII.6 strain GZ2010-L96 detected in China was extensively characterized, and phylogenetic analyses of GII.6 NoVs based on the capsid proteins may reveal a different evolution process from the predominant genotype GII.4. [ABSTRACT FROM AUTHOR]

• Non-epidemic norovirus genotype GII.8 virus-like particles (VLP) were expressed using the baculovirus-insect cell system. • Binding of GII.8 VLP to tissue blood group antigens (HBGAs) was observed. • The cross-reactivity of GII.8 VLP antiserum with other 27 (9 GI genotypes and 18 GII genotypes) capsid protein was evaluated. • Norovirus GII.8 strain antiserum had no cross-blocking effect on the binding of epidemic genotypes (GII.4, GII.17) to HBGA. • The existing vaccine candidate strain GI.1 antiserum and GII.4 antiserum had no cross-blocking effect on the binding of NoV GII.8 to HBGA. Noroviruses are causative agents of acute nonbacterial gastroenteritis epidemics worldwide. There are various genotypes, among which the non-epidemic genotype GII.8 can cause norovirus outbreaks. We previously demonstrated that the immunogenicity of GII.8 differed from that of epidemic variants. This study aimed to comprehensively compare the receptor profile and immunogenicity of the GII.8 variant with those of the epidemic variants. Using the baculovirus-insect cell expression system, we observed that recombinant capsid protein VP1 of the norovirus GII.8 GZ2017-L601 strain formed virus-like particles (VLPs) with a diameter of approximately 30 nm, as evidenced by transmission electron microscopy analysis. The GII.8 VLPs showed weak or moderate binding with all secretor histo-blood group antigens (HBGAs), but not with non-secretors, as evidenced by the HBGA-VLP binding test. The GII.8 VLP antiserum obtained from immunized BALB/c mice was tested for cross-reactivity with other norovirus genotypes (n = 28). The results showed that this antiserum demonstrated moderate cross-reactivity with GI.1, GII.3, and GII.15; however, no cross-reactivity with the epidemic variants of GII.2, GII.4, and GII.17 was observed. Additionally, the blocking-antibody activity of GII.8 antisera against GII.4 VLP-HBGAs and GII.17 VLP-HBGAs interactions and the cross-blocking of GII.8 VLP-HBGAs interactions by GI.1 and GII.4 antisera were evaluated using the HBGAs-VLP blocking test. However, no cross-blocking effect was observed. In summary, the characterization of norovirus GII.8 VLPs and derived antisera revealed that the GII.8 immunogenicity differed from that of epidemic variants. [ABSTRACT FROM AUTHOR]