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1
Aw, Tiong Gim, Karina Yew-Hoong Gin, Lynette Lin Ean Oon, Eileen Xueqin Chen, and Chee Hoe Woo. "Prevalence and Genotypes of Human Noroviruses in Tropical Urban Surface Waters and Clinical Samples in Singapore." Applied and Environmental Microbiology 75, no. 15 (June 12, 2009): 4984–92. http://dx.doi.org/10.1128/aem.00489-09.
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ABSTRACT The prevalence and genotypes of norovirus genogroup I (GI) and GII in tropical urban catchment waters and an estuarine bay were studied. A comparative analysis was performed with environmental isolates of noroviruses and concurrently identified clinical isolates in Singapore during gastroenteritis outbreaks between August 2006 to January 2007. Noroviruses in environmental water samples were concentrated by using ultrafiltration techniques and then analyzed by reverse transcription-seminested PCR assay targeting the partial capsid region of noroviruses and DNA sequencing. Among the 60 water samples collected, noroviruses were detected in 43 (71.7%) of these samples. Of these 43 norovirus-positive samples, the coexistence of both GI and GII strains was identified in 23 (53.5%) water samples. The phylogenetic analysis revealed multiple genotypes of noroviruses GI and GII in environmental water samples. GI and GII strains were clustered into seven and nine (including two unclassified) genotypes, respectively. The major norovirus genotypes in environmental water samples were GI/2 and GI/4 and GII/4. Genotyping of the 21 norovirus-positive clinical samples showed that all of the strains belonged to the GII/4 cluster. The environmental and clinical norovirus GII/4 isolates showed high levels of nucleotide sequence identity to each other and to the novel GII/4 variant associated with global epidemics of gastroenteritis during 2006. This study suggests the emergence and circulation of multiple novel norovirus GI and GII genotypes in water environments. Further comprehensive surveillance of water environments for noroviruses and routine clinical reporting is warranted.
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Lee, Sung-Geun, Weon-Hwa Jheong, Chang-Il Suh, Sang-Hyun Kim, Joong-Bok Lee, Yong-Seok Jeong, GwangPyo Ko, Kyung Lib Jang, Gyu-Cheol Lee, and Soon-Young Paik. "Nationwide Groundwater Surveillance of Noroviruses in South Korea, 2008." Applied and Environmental Microbiology 77, no. 4 (December 23, 2010): 1466–74. http://dx.doi.org/10.1128/aem.01996-10.
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ABSTRACTTo inspect the norovirus contamination of groundwater in South Korea, a nationwide study was performed in the summer (June to August) and winter (October to December) of 2008. Three-hundred sites designated by the government ministry were inspected. Water samples were collected for analysis of water quality, microorganism content, and viral content. Water quality was assessed by temperature, pH, turbidity, residual chlorine, and nitrite nitrogen content. Microorganism contents were analyzed bacteria, total coliforms,Escherichia coli, and bacteriophage. Virus analyses included panenterovirus and norovirus. Two primer sets were used for the detection of norovirus genotypes GI and GII, respectively. Of 300 samples, 65 (21.7%) were norovirus positive in the summer and in 52 (17.3%) were norovirus positive in the winter. The genogroup GI noroviruses that were identified were GI-1, GI-2, GI-3, GI-4, GI-5, GI-6, and GI-8 genotypes; those in the GII genogroup were GII-4 and GII-Yuri genotypes. The analytic data showed correlative relationships between the norovirus detection rate and the following parameters: water temperature and turbidity in physical-chemical parameters and somatic phage in microbial parameters. It is necessary to periodically monitor waterborne viruses that frequently cause epidemic food poisoning in South Korea for better public health and sanitary conditions.
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Munjita, Samuel Munalula. "Current Status of Norovirus Infections in Children in Sub-Saharan Africa." Journal of Tropical Medicine 2015 (2015): 1–7. http://dx.doi.org/10.1155/2015/309648.
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Noroviruses are a leading cause of acute sporadic gastroenteritis worldwide. In Sub-Saharan Africa, information regarding norovirus infections in children is scarce. A systematic review of studies performed between 1993 and June 2015 was conducted to establish the genotypic distribution and prevalence of norovirus infections in children (≤17) in Sub-Saharan Africa. Analysis of data from 19 studies involving 8,399 samples from children with symptomatic and nonsymptomatic gastroenteritis revealed prevalence of 12.6% (range 4.6% to 32.4%). The prevalence of norovirus infections was higher in symptomatic children (14.2%) than asymptomatic children (9.2%). Genogroup II (GII) was the most prevalent genogroup accounting for 76.4% of all the reported norovirus infections. The rest of the infections were GI (21.7%) and GI/GII (1.9%). The most common genotypes were GII.4 (65.2%), GI.7 (33.3%), and GI.3 (21.3%). These statistics were calculated from studies carried out in 12 out of 48 Sub-Saharan African countries. Therefore, more studies involving several countries are required to determine fully the epidemiology of noroviruses and their contribution to childhood diarrhoea in Sub-Saharan Africa.
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Santiso-Bellón, Cristina, Walter Randazzo, Alba Pérez-Cataluña, Susana Vila-Vicent, Roberto Gozalbo-Rovira, Carlos Muñoz, Javier Buesa, Gloria Sanchez, and Jesús Rodríguez Díaz. "Epidemiological Surveillance of Norovirus and Rotavirus in Sewage (2016–2017) in Valencia (Spain)." Microorganisms 8, no. 3 (March 24, 2020): 458. http://dx.doi.org/10.3390/microorganisms8030458.
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The aim of the present study was to perform the molecular epidemiology of rotaviruses and noroviruses detected in sewage samples from a large wastewater facility from the city of Valencia, Spain. A total of 46 sewage samples were collected over a one-year period (September 2016 to September 2017). Norovirus and rotavirus were detected and quantified by RT-qPCR, genotyped by semi-nested RT-PCR and further characterized by sequencing and phylogenetic analyses. Noroviruses and rotaviruses were widely distributed in sewage samples (69.6% for norovirus GI, 76.0% norovirus GII, and 71.7% rotaviruses) and viral loads varied from 4.33 to 5.75 log PCRU/L for norovirus GI, 4.69 to 6.95 log PCRU/L for norovirus GII, and 4.08 to 6.92 log PCRU/L for rotavirus. Overall, 87.5% (28/32) of GI noroviruses could not be genotyped, 6.25% (2/32) of the samples contained GI.2 genotype, and another 6.25% (2/32) were positive for GI.4 genotype. The most common genotype of GII noroviruses was GII.2 (40%, 14/35), followed by GII.6 (8.6%, 3/35) and GII.17 (5.7%, 2/35) while the remaining GII strains could not be typed (45.7%, 16/35). Rotavirus VP4 genotype P[8] was the only one found in 19 out of 33 rotavirus-positive samples (57.7%). G2 was the most prevalent rotavirus VP7 genotype (15.2%, 5/33) followed by G3, G9, and G12, with two positive samples for each genotype (6.1%, 2/33). In one sample both G1 and G2 genotypes were detected simultaneously (3%). The results presented here show that the surveillance of noroviruses and rotaviruses in sewage is useful for the study of their transmission in the population and their molecular epidemiology.
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Kirby, Amy E., Yvonne Kienast, Wanzhe Zhu, Jerusha Barton, Emeli Anderson, Melissa Sizemore, Jan Vinje, and Christine L. Moe. "Norovirus Seroprevalence among Adults in the United States: Analysis of NHANES Serum Specimens from 1999–2000 and 2003–2004." Viruses 12, no. 2 (February 5, 2020): 179. http://dx.doi.org/10.3390/v12020179.
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Norovirus is the most common cause of epidemic and endemic acute gastroenteritis. However, national estimates of the infection burden are challenging. This study used a nationally representative serum bank to estimate the seroprevalence to five norovirus genotypes including three GII variants: GI.1 Norwalk, GI.4, GII.3, GII.4 US95/96, GII.4 Farmington Hills, GII.4 New Orleans, and GIV.1 in the USA population (aged 16 to 49 years). Changes in seroprevalence to the three norovirus GII.4 variants between 1999 and 2000, as well as 2003 and 2004, were measured to examine the role of population immunity in the emergence of pandemic GII.4 noroviruses. The overall population-adjusted seroprevalence to any norovirus was 90.0% (1999 to 2000) and 95.9% (2003 to 2004). Seroprevalence was highest to GI.1 Norwalk, GII.3, and the three GII.4 noroviruses. Seroprevalence to GII.4 Farmington Hills increased significantly between the 1999 and 2000, as well as the 2003 and 2004, study cycles, consistent with the emergence of this pandemic strain. Seroprevalence to GII.4 New Orleans also increased over time, but to a lesser degree. Antibodies against the GIV.1 norovirus were consistently detected (population-adjusted seroprevalence 19.1% to 25.9%), with rates increasing with age. This study confirms the high burden of norovirus infection in US adults, with most adults having multiple norovirus infections over their lifetime.
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Sarmento, Sylvia Kahwage, Juliana da Silva Ribeiro de Andrade, Marize Pereira Miagostovich, and Tulio Machado Fumian. "Virological and Epidemiological Features of Norovirus Infections in Brazil, 2017–2018." Viruses 13, no. 9 (August 30, 2021): 1724. http://dx.doi.org/10.3390/v13091724.
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Noroviruses are considered an important cause of acute gastroenteritis (AGE) across all age groups. Here, we investigated the incidence of norovirus, genotypes circulation, and norovirus shedding in AGE stool samples from outpatients in Brazil. During a two-year period, 1546 AGE stool samples from ten Brazilian states were analyzed by RT-qPCR to detect and quantify GI and GII noroviruses. Positive samples were genotyped by dual sequencing using the ORF1/2 junction region. Overall, we detected norovirus in 32.1% of samples, with a massive predominance of GII viruses (89.1%). We also observed a significant difference between the median viral load of norovirus GI (3.4×105 GC/g of stool) and GII (1.9×107 GC/g). The most affected age group was children aged between 6 and 24 m old, and norovirus infection was detected throughout the year without marked seasonality. Phylogenetic analysis of partial RdRp and VP1 regions identified six and 11 genotype combinations of GI and GII, respectively. GII.4 Sydney[P16] was by far the predominant genotype (47.6%), followed by GII.2[P16], GII.4 Sydney[P31], and GII.6[P7]. We detected, for the first time in Brazil, the intergenogroup recombinant genotype GIX.1[GII.P15]. Our study contributes to the knowledge of norovirus genotypes circulation at the national level, reinforcing the importance of molecular surveillance programs for future vaccine designs.
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Jung, James, Timothy Grant, Dennis R. Thomas, Chris W. Diehnelt, Nikolaus Grigorieff, and Leemor Joshua-Tor. "High-resolution cryo-EM structures of outbreak strain human norovirus shells reveal size variations." Proceedings of the National Academy of Sciences 116, no. 26 (June 10, 2019): 12828–32. http://dx.doi.org/10.1073/pnas.1903562116.
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Noroviruses are a leading cause of foodborne illnesses worldwide. Although GII.4 strains have been responsible for most norovirus outbreaks, the assembled virus shell structures have been available in detail for only a single strain (GI.1). We present high-resolution (2.6- to 4.1-Å) cryoelectron microscopy (cryo-EM) structures of GII.4, GII.2, GI.7, and GI.1 human norovirus outbreak strain virus-like particles (VLPs). Although norovirus VLPs have been thought to exist in a single-sized assembly, our structures reveal polymorphism between and within genogroups, with small, medium, and large particle sizes observed. Using asymmetric reconstruction, we were able to resolve a Zn2+metal ion adjacent to the coreceptor binding site, which affected the structural stability of the shell. Our structures serve as valuable templates for facilitating vaccine formulations.
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Lindesmith, Lisa C., Eric Donaldson, Juan Leon, Christine L. Moe, Jeffrey A. Frelinger, Robert E. Johnston, David J. Weber, and Ralph S. Baric. "Heterotypic Humoral and Cellular Immune Responses following Norwalk Virus Infection." Journal of Virology 84, no. 4 (December 9, 2009): 1800–1815. http://dx.doi.org/10.1128/jvi.02179-09.
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ABSTRACT Norovirus immunity is poorly understood as the limited data available on protection after infection are often contradictory. In contrast to the more prominent GII noroviruses, GI norovirus infections are less frequent in outbreaks. The GI noroviruses display very complex patterns of heterotypic immune responses following infection, and many individuals are highly susceptible to reinfection. To study the immune responses and mechanisms of GI.1 persistence, we built structural models and recombinant virus-like particles (VLPs) of five GI strains: GI.1-1968, GI.1-2001, GI.2-1999, GI.3-1999, and GI.4-2000. Structural models of four GI genotype capsid P domain dimers suggested that intragenotype structural variation is limited, that the GI binding pocket is mostly preserved between genotypes, and that a conserved, surface-exposed epitope may allow for highly cross-reactive immune responses. GI VLPs bound to histo-blood group antigens (HBGAs) including fucose, Lewis, and A antigens. Volunteers infected with GI.1-1968 (n = 10) had significant increases between prechallenge and convalescent reactive IgG for all five GI VLPs measured by enzyme immunoassay. Potential cross-neutralization of GI VLPs was demonstrated by convalescent-phase serum cross-blockade of GI VLP-HBGA interaction. Although group responses were significant for all GI VLPs, each individual volunteer demonstrated a unique VLP blockade pattern. Further, peripheral blood mononuclear cells (PBMCs) were stimulated with each of the VLPs, and secretion of gamma interferon (IFN-γ) was measured. As seen with blockade responses, IFN-γ secretion responses differed by individual. Sixty percent responded to at least one GI VLP, with only two volunteers responding to GI.1 VLP. Importantly, four of five individuals with sufficient PBMCs for cross-reactivity studies responded more robustly to other GI VLPs. These data suggest that preexposure history and deceptive imprinting may complicate PBMC and B-cell immune responses in some GI.1-1968-challenged individuals and highlight a potential complication in the design of efficacious norovirus vaccines.
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Hesse, Shayla, Frederick H. Neill, Mary K. Estes, Sreejesh Shanker, B. V. Venkataram Prasad, Jennifer Ferreira, and Robert L. Atmar. "Serological Responses to a Norovirus Nonstructural Fusion Protein after Vaccination and Infection." Clinical and Vaccine Immunology 23, no. 2 (December 9, 2015): 181–83. http://dx.doi.org/10.1128/cvi.00595-15.
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ABSTRACTThe performance of an assay to detect antibodies to a norovirus nonstructural fusion protein, designated VPR and consisting of three proteins (GI.1 virus protein genome-linked [VPg], a virus protease, and an RNA-dependent RNA polymerase), was evaluated. The assay sensitivity and specificity were 74.5% and >95%, respectively, for identifying GI.1 norovirus infection among persons who received either a monovalent GI.1 norovirus virus-like particle (VLP) vaccine or placebo by the intranasal route followed by an oral live GI.1 norovirus challenge.
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Cannon, Jennifer L., Leslie Barclay, Nikail R. Collins, Mary E. Wikswo, Christina J. Castro, Laura Cristal Magaña, Nicole Gregoricus, Rachel L. Marine, Preeti Chhabra, and Jan Vinjé. "Genetic and Epidemiologic Trends of Norovirus Outbreaks in the United States from 2013 to 2016 Demonstrated Emergence of Novel GII.4 Recombinant Viruses." Journal of Clinical Microbiology 55, no. 7 (May 10, 2017): 2208–21. http://dx.doi.org/10.1128/jcm.00455-17.
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ABSTRACT Noroviruses are the most frequent cause of epidemic acute gastroenteritis in the United States. Between September 2013 and August 2016, 2,715 genotyped norovirus outbreaks were submitted to CaliciNet. GII.4 Sydney viruses caused 58% of the outbreaks during these years. A GII.4 Sydney virus with a novel GII.P16 polymerase emerged in November 2015, causing 60% of all GII.4 outbreaks in the 2015-2016 season. Several genotypes detected were associated with more than one polymerase type, including GI.3, GII.2, GII.3, GII.4 Sydney, GII.13, and GII.17, four of which harbored GII.P16 polymerases. GII.P16 polymerase sequences associated with GII.2 and GII.4 Sydney viruses were nearly identical, suggesting common ancestry. Other common genotypes, each causing 5 to 17% of outbreaks in a season, included GI.3, GI.5, GII.2, GII.3, GII.6, GII.13, and GII.17 Kawasaki 308. Acquisition of alternative RNA polymerases by recombination is an important mechanism for norovirus evolution and a phenomenon that was shown to occur more frequently than previously recognized in the United States. Continued molecular surveillance of noroviruses, including typing of both polymerase and capsid genes, is important for monitoring emerging strains in our continued efforts to reduce the overall burden of norovirus disease.
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da Silva, Allegra Kyria, Jean-Claude Le Saux, Sylvain Parnaudeau, Monique Pommepuy, Menachem Elimelech, and Françoise S. Le Guyader. "Evaluation of Removal of Noroviruses during Wastewater Treatment, Using Real-Time Reverse Transcription-PCR: Different Behaviors of Genogroups I and II." Applied and Environmental Microbiology 73, no. 24 (October 12, 2007): 7891–97. http://dx.doi.org/10.1128/aem.01428-07.
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ABSTRACT Noroviruses, an important cause of gastroenteritis, are excreted by infected individuals and are therefore present in wastewater. We quantified norovirus genogroup I (GI) and GII in wastewater at different locations in France and evaluated removal by a range of treatment types, including basic (waste stabilization pond), current industry standard (activated sludge), and state-of-the-art (submerged membrane bioreactor) treatments. Noroviruses were quantified using real-time reverse transcription-PCR (rRT-PCR). Mengovirus was used as a virus extraction control, and internal controls were used to verify the level of GI and GII rRT-PCR inhibition. A total of 161 (81 influent and 79 effluent) samples were examined; GI and GII were detected in 43 and 88% of the influent samples, respectively, and in 24 and 14% of the effluent samples, respectively. Physicians in France report far more cases of GII than GI during outbreaks; thus, the frequent presence of GI was unexpected. The GI influent concentrations were more variable, the peak GI influent concentrations were higher than the peak GII influent concentrations at all four sites (up to 1 × 109 and 6 × 107 genome copies/liter, respectively), and the average positive influent concentrations of GI were higher than the average positive influent concentrations of GII. The maximum effluent breakthrough concentrations were 6 × 106 and 3 × 106 genome copies/liter for GI and GII, respectively, indicating that the four treatment systems studied decreased the norovirus contamination load in receiving waters.
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BRUGGINK, L. D., N. L. DUNBAR, and J. A. MARSHALL. "Norovirus genotype diversity associated with gastroenteritis outbreaks in aged-care facilities." Epidemiology and Infection 143, no. 14 (February 27, 2015): 3064–68. http://dx.doi.org/10.1017/s095026881500031x.
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SUMMARYNoroviruses are a major cause of gastroenteritis. 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 residential aged-care facilities in Victoria, Australia over one year (2013) and documented the (capsid) norovirus genotypes associated with these outbreaks. It was found that 65·0% of 206 outbreaks tested were associated with norovirus infection, thereby showing norovirus to be the major cause of viral gastroenteritis in residential aged-care facilities. Fifteen capsid (open reading frame 2) genotypes were identified as follows: GI.2 (0·9%), GI.3 (1·8%), GI.4 (3·7%), GI.6 (0·9%), GI.7 (0·9%), GI.8 (0·9%), GII.1 (0·9%), GII.2 (0·9%), GII.3 (1·8%), GII.4 (2009-like) (0·9%), GII.4 (2012) (48·6%), GII.4 (2012-like) (16·5%), GII.4 (unknown) (9·2%), GII.5 (2·8%), GII.6 (0·9%), GII.7 (0·9%), GII.13 (6·4%) and an as yet unclassified GII genotype (0·9%). Although GII.4 was the most common norovirus capsid genotype detected, the great diversity of norovirus genotypes in the elderly indicates vaccination strategies for this demographic are not straightforward.
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Mans. "Norovirus Infections and Disease in Lower‐Middleand Low‐Income Countries, 1997–2018." Viruses 11, no. 4 (April 10, 2019): 341. http://dx.doi.org/10.3390/v11040341.
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Noroviruses are a major cause of viral gastroenteritis. The burden of the norovirus in lowresourcesettings is not well‐established due to limited data. This study reviews the norovirusprevalence, epidemiology, and genotype diversity in lower‐middle‐income countries (LMIC) andin low‐income countries (LIC). PubMed was searched up to 14 January 2019 for norovirus studiesfrom all LIC and LMIC (World Bank Classification). Studies that tested gastroenteritis cases and/orasymptomatic controls for norovirus by reverse transcription‐polymerase chain reaction (RT‐PCR)were included. Sixty‐four studies, the majority on children <5 years of age, were identified, and 14%(95% confidence interval; CI 14–15, 5158/36,288) of the gastroenteritis patients and 8% (95% CI 7–9,423/5310) of healthy controls tested positive for norovirus. In LMIC, norovirus was detected in 15%(95% CI 15–16) of cases and 8% (95% CI 8–10) of healthy controls. In LIC, 11% (95% CI 10–12) ofsymptomatic cases and 9% (95% CI 8–10) of asymptomatic controls were norovirus positive.Norovirus genogroup II predominated overall. GII.4 was the predominant genotype in all settings,followed by GII.3 and GII.6. The most prevalent GI strain was GI.3. Norovirus causes a significantamount of gastroenteritis in low‐resource countries, albeit with high levels of asymptomaticinfection in LIC and a high prevalence of coinfections
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Zhang, Lifang, Yanxia Peng, Na Jiang, Lei Shi, Jieping Lin, Ping Wu, and Qingjun Pan. "Preparation and Evaluation of Combined Detection of Norovirus GI and GII: An Innovative Fluorescent Particles Test Strip." BioMed Research International 2018 (2018): 1–5. http://dx.doi.org/10.1155/2018/7862467.
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This study was designed to prepare and evaluate the sensitivity and specificity of a Norovirus GI and GII fluorescent particles combined detection test strip method. Using selected chromatographic materials and antibodies specific to Norovirus GI and GII, the Norovirus GI and GII fluorescent particles combined detection test strip (tested method) was prepared as a conventional double antibody sandwich. The samples assayed included cultured rotavirus and 465 specimens from patients with symptoms of gastrointestinal infection. Norovirus was detected using the tested method and a reference method (CerTest Norovirus GI-GII test card). The results indicated that the sensitivity of the tested method was 4 (for GI detection) or 8 times (for GII detection) greater than the reference method. Neither of the two methods cross-reacted with rotavirus and so on. For specimens, 29 were found to be negative by the reference method and positive by the tested method, and 8 were found to be negative by the tested method and positive by the reference method. Furthermore, a retesting of these samples by qPCR showed that 28 of the 29 were positive, and 3 of the 8 were positive. In summary, the Norovirus GI and GII fluorescent particles combined detection test strip was successfully prepared and had good detection performance.
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Chiu, Shu-Chun, Jia-Kai Hsu, Szu-Chieh Hu, Ching-Yi Wu, Ying-Chin Wang, and Jih-Hui Lin. "Molecular Epidemiology of GI.3 Norovirus Outbreaks from Acute Gastroenteritis Surveillance System in Taiwan, 2015–2019." BioMed Research International 2020 (February 13, 2020): 1–9. http://dx.doi.org/10.1155/2020/4707538.
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Norovirus is the leading cause of food-borne disease outbreaks. We conducted this study to examine the incidence and molecular characteristics of norovirus genogroup I infections from acute gastroenteritis outbreaks in Taiwan. Between January 2015 and June 2019, 2121 acute gastroenteritis clusters were reported to Taiwan CDC, of which 351 (16.5%) clusters were positive for NoV GI, and GI.3 was the most prevalent (36.8%) during the study period. The GI.3 infections were significantly higher than non-GI.3 infections in the age groups of 0–5 and 6–18 years. The phylogenetic analysis of the MCC tree revealed that VP1 genes were divided into 3 groups: the GI.P3-GI.3 strains in Taiwan were genetically close to Japan and the GI.Pd-GI.3 strains were segregated into 2 other groups which were genetically closely related to China. In addition, 7 GI.Pd-GI.3 recombinants were identified circulating in Taiwan between 2018 and 2019, and the prevalence of GI.Pd-GI.3 should be monitored to assess whether this could become the new predominant strains in neighboring Asian countries or other parts of the world. Both GI.P3-GI.3 and GI.Pd-GI.3 strains cocirculate, the recombination among these two lineages occurs frequently, contributing to the genetic diversity and multiple occurrences of different norovirus lineages, and their rapid evolution makes future control more difficult. Continued surveillance and timely interventions are critical to understand the complexity of norovirus gene variation and to monitor the new emerging norovirus strains.
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Gonzalez, Mark D., L. Claire Langley, Blake W. Buchan, Matthew L. Faron, Melanie Maier, Kate Templeton, Kimberly Walker, et al. "Multicenter Evaluation of the Xpert Norovirus Assay for Detection of Norovirus Genogroups I and II in Fecal Specimens." Journal of Clinical Microbiology 54, no. 1 (November 11, 2015): 142–47. http://dx.doi.org/10.1128/jcm.02361-15.
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Norovirus is the most common cause of sporadic gastroenteritis and outbreaks worldwide. The rapid identification of norovirus has important implications for infection prevention measures and may reduce the need for additional diagnostic testing. The Xpert Norovirus assay recently received FDA clearance for the detection and differentiation of norovirus genogroups I and II (GI and GII), which account for the vast majority of infections. In this study, we evaluated the performance of the Xpert Norovirus assay with both fresh, prospectively collected (n= 914) and frozen, archived (n= 489) fecal specimens. A Centers for Disease Control and Prevention (CDC) composite reference method was used as the gold standard for comparison. For both prospective and frozen specimens, the Xpert Norovirus assay showed positive percent agreement (PPA) and negative percent agreement (NPA) values of 98.3% and 98.1% for GI and of 99.4% and 98.2% for GII, respectively. Norovirus prevalence in the prospective specimens (collected from March to May of 2014) was 9.9% (n= 90), with the majority of positives caused by genogroup II (82%,n= 74). The positive predictive value (PPV) of the Xpert Norovirus assay was 75% for GI-positive specimens, whereas it was 86.5% for GII-positive specimens. The negative predictive values (NPV) for GI and GII were 100% and 99.9%, respectively.
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Yuki, Yoshikazu, Shiho Kurokawa, Shintaro Sato, Ai Sasou, Naomi Matsumoto, Akio Suzuki, Naomi Sakon, et al. "A Heterodimeric Antibody Fragment for Passive Immunotherapy Against Norovirus Infection." Journal of Infectious Diseases 222, no. 3 (March 25, 2020): 470–78. http://dx.doi.org/10.1093/infdis/jiaa115.
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Abstract Human noroviruses cause an estimated 685 million infections and 200 000 deaths annually worldwide. Although vaccines against GII.4 and GI.1 genotypes are under development, no information is available regarding vaccines or monoclonal antibodies to other noroviral genotypes. Here, we developed 2 variable-domain llama heavy-chain antibody fragment (VHHs) clones, 7C6 and 1E4, against GII.4 and GII.17 human noroviruses, respectively. Although 7C6 cross-reacted with virus-like particles (VLPs) of GII.17, GII.6, GII.3, and GII.4, it neutralized only GII.4 norovirus. In contrast, 1E4 reacted with and neutralized only GII.17 VLPs. Both VHHs blocked VLP binding to human induced pluripotent stem cell-derived intestinal epithelial cells and carbohydrate attachment factors. Using these 2 VHHs, we produced a heterodimeric VHH fragment that neutralized both GII.4 and GII.17 noroviruses. Because VHH fragments are heat- and acid-stable recombinant monoclonal antibodies, the heterodimer likely will be useful for oral immunotherapy and prophylaxis against GII.4 and GII.17 noroviruses in young, elderly, or immunocompromised persons.
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Rossouw, Esmari, Marieke Brauer, Pieter Meyer, Nicolette M. du Plessis, Theunis Avenant, and Janet Mans. "Virus Etiology, Diversity and Clinical Characteristics in South African Children Hospitalised with Gastroenteritis." Viruses 13, no. 2 (January 30, 2021): 215. http://dx.doi.org/10.3390/v13020215.
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Background: Viral gastroenteritis remains a major cause of hospitalisation in young children. This study aimed to determine the distribution and diversity of enteric viruses in children ≤5 years, hospitalised with gastroenteritis at Kalafong Provincial Tertiary Hospital, Pretoria, South Africa, between July 2016 and December 2017. Methods: Stool specimens (n = 205) were screened for norovirus GI and GII, rotavirus, sapovirus, astrovirus and adenovirus by multiplex RT-PCR. HIV exposure and FUT2 secretor status were evaluated. Secretor status was determined by FUT2 genotyping. Results: At least one gastroenteritis virus was detected in 47% (96/205) of children. Rotavirus predominated (46/205), followed by norovirus (32/205), adenovirus (15/205), sapovirus (9/205) and astrovirus (3/205). Norovirus genotypes GI.3, GII.2, GII.3, GII.4, GII.7, GII.12, GII.21, and rotavirus strains G1P[8], G2P[4], G2P[6], G3P[4], G3P[8], G8P[4], G8P[6], G9P[6], G9P[8] and sapovirus genotypes GI.1, GI.2, GII.1, GII.4, GII.8 were detected; norovirus GII.4[P31] and rotavirus G3P[4] predominated. Asymptomatic norovirus infection (GI.3, GI.7, GII.4, GII.6, GII.13) was detected in 22% of 46 six-week follow up stools. HIV exposure (30%) was not associated with more frequent or severe viral gastroenteritis hospitalisations compared to unexposed children. Rotavirus preferentially infected secretor children (p = 0.143) and norovirus infected 78% secretors and 22% non-secretors. Conclusion: Rotavirus was still the leading cause of gastroenteritis hospitalisations, but norovirus caused more severe symptoms.
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Montazeri, Naim, Dorothee Goettert, Eric C. Achberger, Crystal N. Johnson, Witoon Prinyawiwatkul, and Marlene E. Janes. "Pathogenic Enteric Viruses and Microbial Indicators during Secondary Treatment of Municipal Wastewater." Applied and Environmental Microbiology 81, no. 18 (July 10, 2015): 6436–45. http://dx.doi.org/10.1128/aem.01218-15.
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ABSTRACTPathogenic enteric viruses are responsible for a wide range of infections in humans, with diverse symptoms. Raw and partially treated wastewaters are major sources of environmental contamination with enteric viruses. We monitored a municipal secondary wastewater treatment plant (New Orleans, LA) on a monthly basis for norovirus (NoV) GI and GII and enterovirus serotypes using multiplex reverse transcription-quantitative PCR (RT-qPCR) and microbial indicators of fecal contamination using standard plating methods. Densities of indicator bacteria (enterococci, fecal coliforms, andEscherichia coli) did not show monthly or seasonal patterns. Norovirus GII was more abundant than GI and, along with enterovirus serotypes, increased in influent during fall and spring. The highest NoV GI density in influent was in the fall, reaching an average of 4.0 log10genomic copies/100 ml. Norovirus GI removal (0.95 log10) was lower than that for GII, enterovirus serotypes, and male-specific coliphages (1.48 log10) or for indicator bacteria (4.36 log10), suggesting higher resistance of viruses to treatment. Male-specific coliphages correlated with NoV GII densities in influent and effluent (r= 0.48 and 0.76, respectively) and monthly removal, indicating that male-specific coliphages can be more reliable than indicator bacteria to monitor norovirus GII load and microbial removal. Dominant norovirus genotypes were classified into three GI genotypes (GI.1, GI.3, and GI.4) and four GII genotypes (GII.3, GII.4, GII.13, and GII.21), dominated by GI.1 and GII.4 strains. Some of the seasonal and temporal patterns we observed in the pathogenic enteric viruses were different from those of epidemiological observations.
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Prystajecky, Natalie, Fiona SL Brinkman, Brian Auk, Judith L. Isaac-Renton, and Patrick Tang. "Personalized Genetic Testing and Norovirus Susceptibility." Canadian Journal of Infectious Diseases and Medical Microbiology 25, no. 4 (2014): 222–24. http://dx.doi.org/10.1155/2014/708579.
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BACKGROUND: The availability of direct-to-consumer personalized genetic testing has enabled the public to access and interpret their own genetic information. Various genetic traits can be determined including resistance to norovirus through a nonsense mutation (G428A) in theFUT2gene. Although this trait is believed to confer resistance to the most dominant norovirus genotype (GII.4), the spectrum of resistance to other norovirus strains is unknown. The present report describes a cluster of symptomatic norovirus GI.6 infection in a family identified to have norovirus resistance through personalized genetic testing.CASE PRESENTATION: In January 2013, four members of a family determined by a direct-to-consumer genetic test to be homozygous for the norovirus resistance trait (A/A genotype for single nucleotide polymorphism rs601338) developed symptoms consistent with acute viral gastroenteritis. Stool and vomitus samples were submitted for enteric viral pathogen testing. Samples were positive for norovirus GI.6 in three of the four cases.CONCLUSIONS: The present report is the first to describe norovirus GI.6 infection in patients with the G428A nonsense mutation inFUT2; this cluster of cases suggests that the G428A mutation inFUT2may not confer resistance to norovirus GI.6. Direct-to-consumer genetic testing is empowering members of the public to identify novel associations with their genetic traits. Expert consultation is important for the interpretation of personalized genetic test results, and follow-up laboratory testing can confirm any potentially novel associations.
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Schilling-Loeffler, Katja, Rachel Rodriguez, and Jacquelina Williams-Woods. "Target Affinity and Structural Analysis for a Selection of Norovirus Aptamers." International Journal of Molecular Sciences 22, no. 16 (August 18, 2021): 8868. http://dx.doi.org/10.3390/ijms22168868.
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Aptamers, single-stranded oligonucleotides that specifically bind a molecule with high affinity, are used as ligands in analytical and therapeutic applications. For the foodborne pathogen norovirus, multiple aptamers exist but have not been thoroughly characterized. Consequently, there is little research on aptamer-mediated assay development. This study characterized seven previously described norovirus aptamers for target affinity, structure, and potential use in extraction and detection assays. Norovirus-aptamer affinities were determined by filter retention assays using norovirus genotype (G) I.1, GI.7, GII.3, GII.4 New Orleans and GII.4 Sydney virus-like particles. Of the seven aptamers characterized, equilibrium dissociation constants for GI.7, GII.3, GII.4 New Orleans and GII.4 Sydney ranged from 71 ± 38 to 1777 ± 1021 nM. Four aptamers exhibited affinity to norovirus GII.4 strains; three aptamers additionally exhibited affinity toward GII.3 and GI.7. Aptamer affinity towards GI.1 was not observed. Aptamer structure analysis by circular dichroism (CD) spectroscopy showed that six aptamers exhibit B-DNA structure, and one aptamer displays parallel/antiparallel G-quadruplex hybrid structure. CD studies also showed that biotinylated aptamer structures were unchanged from non-biotinylated aptamers. Finally, norovirus aptamer assay feasibility was demonstrated in dot-blot and pull-down assays. This characterization of existing aptamers provides a knowledge base for future aptamer-based norovirus detection and extraction assay development and aptamer modification.
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Ritchie, Arabela, Armando Hung, and Muriel Gómez-Sánchez. "Detección de Norovirus GI Y GII en muestras de agua del Río Piura mediante la técnica de RT-PCR en tiempo real." Salud y Tecnología Veterinaria 6, no. 2 (February 15, 2019): 47. http://dx.doi.org/10.20453/stv.v6i2.3458.
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Los Norovirus (NoV) son una de las causas más frecuentes de infecciones gastrointestinales agudas en humanos y un problema de salud importante en países en desarrollo. El objetivo del estudio fue detectar la presencia del Norovirus GI y GII en muestras de agua del Rio Piura mediante la concentración del virus en agua y RT-PCR en tiempo real. Para ello se recolectaron muestras de agua en botellas estériles en tres (3) puntos determinados del río, cada 15 días, desde mayo hasta Setiembre del 2013. Se obtuvo 20.8% (5/24) de muestras positivas para Norovirus GI mediante la técnica de RT-PCR en tiempo real. Ninguna de las muestras dio positivo a Norovirus GII. Este es el primer reporte de Norovirus GI en muestras de agua del Rio Piura. Los métodos propuestos podrían ser utilizados como herramienta para la investigación de patógenos en aguas del Rio Piura y la Bahía de Sechura.
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Wolf, Sandro, Wendy M. Williamson, Joanne Hewitt, Malet Rivera-Aban, Susan Lin, Andrew Ball, Paula Scholes, and Gail E. Greening. "Sensitive Multiplex Real-Time Reverse Transcription-PCR Assay for the Detection of Human and Animal Noroviruses in Clinical and Environmental Samples." Applied and Environmental Microbiology 73, no. 17 (July 6, 2007): 5464–70. http://dx.doi.org/10.1128/aem.00572-07.
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ABSTRACT In this study, we developed a triplex real-time reverse transcription-PCR (RT-PCR)-based method that detects and distinguishs between noroviruses belonging to genogroups I, II, and III and that targets the junction between the regions of open reading frame 1 (ORF1) and ORF2. This is the first assay to include all three genogroups and the first real-time RT-PCR-based method developed for the detection of bovine noroviruses. The assay was shown to be broadly reactive against a wide spectrum of norovirus genotypes, including GI/1 through GI/7, GII/1 through GII/8, GII/10, GII/12, and GII/17, in different matrices (including fecal specimens, treated and raw sewage, source water, and treated drinking water). The assay is highly sensitive, detecting low copy numbers of plasmids that carry the target sequence. A new bovine norovirus, Bo/NLV/Norsewood/2006/NZL, was identified by this assay and was further genetically characterized. The results implicate a broad range of possible applications, including clinical diagnostics, tracing of fecal contaminants, and due to its sensitivity and broad reactivity, environmental studies.
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Van Dycke, Jana, Wenhao Dai, Zoe Stylianidou, Jian Li, Arno Cuvry, Emma Roux, Bingqian Li, et al. "A Novel Class of Norovirus Inhibitors Targeting the Viral Protease with Potent Antiviral Activity In Vitro and In Vivo." Viruses 13, no. 9 (September 16, 2021): 1852. http://dx.doi.org/10.3390/v13091852.
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Human noroviruses (HuNoVs) are the most common cause of viral gastroenteritis resulting annually in ~219,000 deaths and a societal cost of ~USD 60 billion, and no antivirals or vaccines are available. Here, we assess the anti-norovirus activity of new peptidomimetic aldehydes related to the protease inhibitor rupintrivir. The early hit compound 4 inhibited the replication of murine norovirus (MNV) and the HuNoV GI.1 replicon in vitro (EC50 ~1 µM) and swiftly cleared the HuNoV GI.1 replicon from the cells. Compound 4 still inhibits the proteolytic activity. We selected a resistant GI.1 replicon, with a mutation (I109V) in a highly conserved region of the viral protease, conferring a low yield of resistance against compound 4 and rupintrivir. After testing new derivatives, compound 10d was the most potent (EC50 nanomolar range). Molecular docking indicated that the aldehyde group of compounds 4 and 10d bind with Cys139 in the HuNoV 3CL protease by a covalent linkage. Finally, compound 10d inhibited the replication of HuNoV GII.4 in infected zebrafish larvae, and PK studies in mice showed an adequate profile.
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Nahar, Shamsun, Mokibul Hassan Afrad, Noorjahan Begum, Feroz Al-Mamun, Azadul Kabir Sarker, Sumon Kumar Das, Abu SG Faruque, et al. "High prevalence of noroviruses among hospitalized diarrheal patients in Bangladesh, 2011." Journal of Infection in Developing Countries 7, no. 11 (November 15, 2013): 892–96. http://dx.doi.org/10.3855/jidc.2944.
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Introduction: Norovirus is not usually investigated in diarrheal patients in Bangladesh which may account for the many cases where no pathogens are identified. Methodology: Stool specimens collected from diarrheal patients from three hospitals in Bangladesh during 2011 were investigated for norovirus RNA using real-time RT-PCR assay with norovirus type specific primers and probes. Results: Of the 257 stool specimens tested, 28.4 % were norovirus positive. GII (71.2%) was the predominant strain followed by GI (20.5%), GI+GII (6.8%) and GIV (1.4%). Half of the norovirus positive stools (n=37) were co-infected with other pathogens. Conclusion: Continued surveillance of norovirus together with other viral and bacterial pathogens in hospitalized gastroenteritis patients as well as in the community will further elucidate the role and burden of different pathogens in diarrheal diseases.
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Chhabra, Preeti, Miranda de Graaf, Gabriel I. Parra, Martin Chi-Wai Chan, Kim Green, Vito Martella, Qiuhong Wang, et al. "Updated classification of norovirus genogroups and genotypes." Journal of General Virology 100, no. 10 (October 1, 2019): 1393–406. http://dx.doi.org/10.1099/jgv.0.001318.
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Noroviruses are genetically diverse RNA viruses associated with acute gastroenteritis in mammalian hosts. Phylogenetically, they can be segregated into different genogroups as well as P (polymerase)-groups and further into genotypes and P-types based on amino acid diversity of the complete VP1 gene and nucleotide diversity of the RNA-dependent RNA polymerase (RdRp) region of ORF1, respectively. In recent years, several new noroviruses have been reported that warrant an update of the existing classification scheme. Using previously described 2× standard deviation (sd) criteria to group sequences into separate clusters, we expanded the number of genogroups to 10 (GI-GX) and the number of genotypes to 49 (9 GI, 27 GII, 3 GIII, 2 GIV, 2 GV, 2 GVI and 1 genotype each for GVII, GVIII, GIX [formerly GII.15] and GX). Viruses for which currently only one sequence is available in public databases were classified into tentative new genogroups (GNA1 and GNA2) and genotypes (GII.NA1, GII.NA2 and GIV.NA1) with their definitive assignment awaiting additional related sequences. Based on nucleotide diversity in the RdRp region, noroviruses can be divided into 60 P-types (14 GI, 37 GII, 2 GIII, 1 GIV, 2 GV, 2 GVI, 1 GVII and 1 GX), 2 tentative P-groups and 14 tentative P-types. Future classification and nomenclature updates will be based on complete genome sequences and will be coordinated and disseminated by the international norovirus classification-working group.
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Maalouf, Haifa, Maha Zakhour, Jacques Le Pendu, Jean-Claude Le Saux, Robert L. Atmar, and Françoise S. Le Guyader. "Distribution in Tissue and Seasonal Variation of Norovirus Genogroup I and II Ligands in Oysters." Applied and Environmental Microbiology 76, no. 16 (June 18, 2010): 5621–30. http://dx.doi.org/10.1128/aem.00148-10.
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ABSTRACT Bivalve molluscan shellfish, such as oysters, filter large volumes of water as part of their feeding activities and are able to accumulate and concentrate different types of pathogens, particularly noroviruses, from fecal human pollution. Based on our previous observation of a specific binding of the Norwalk strain (prototype norovirus genogroup I) to the oyster digestive tract through an A-like carbohydrate structure indistinguishable from human blood group A antigen and on the large diversity between strains in terms of carbohydrate-binding specificities, we evaluated the different ligands implicated in attachment to oysters tissues of strains representative of two main genogroups of human norovirus. The GI.1 and GII.4 strains differed in that the latter recognized a sialic acid-containing ligand, present in all tissues, in addition to the A-like ligand of the digestive tract shared with the GI.1 strain. Furthermore, bioaccumulation experiments using wild-type or mutant GI.1 Viruslike particles showed accumulation in hemocytes largely, but not exclusively, based on interaction with the A-like ligand. Moreover, a seasonal effect on the expression of these ligands was detected, most visibly for the GI.1 strain, with a peak in late winter and spring, a period when GI strains are regularly involved in oyster-related outbreaks. These observations may explain some of the distinct epidemiological features of strains from different genogroups.
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Krumova-Valcheva, G., Z. Mladenova, and Y. Gogov. "Study on norovirus contamination of live bivalve molluscs using real-time PCR." BULGARIAN JOURNAL OF VETERINARY MEDICINE 23, no. 4 (2020): 478–86. http://dx.doi.org/10.15547/bjvm.2019-0008.
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Foodborne and waterborne viruses are a major cause of human morbidity. Of them, noroviruses are recognised as the leading causative agents of sporadic infections and epidemic outbreaks of acute gastroenteritis in humans. Contaminated food products and water are the main source of infection with noroviruses. The infection of bivalve molluscs with human pathogenic viruses occurs by faecal contamination in the production coastal waters. In this study, 47 samples of live bivalve molluscs, including 15 samples of cultivated mussels (Mytilus galloprovincialis) and 32 samples of wild mussels (Tapes decussatus), collected from the Bulgarian and Mediterranean coasts, respectively, were submitted to RT-real-time TaqMan PCR to detect the presence of noroviruses genotype GI and GII. Norovirus genotype GII was found in 11 (23.4%) of all the samples tested. A single mollusc sample (2.1%) was positive for both norovirus genotypes. Our results demonstrated that shellfish intended for sale on the Bulgarian market might pose a potential risk for acquiring norovirus infection. Thus, food safety quality control of shellfish by optimised and standardised methods for detection of foodborne viruses, including noroviruses, should be urgently implemented in Bulgaria.
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Fardah Athiyyah, Alpha, Katsumi Shigemura, Koichi Kitagawa, Nazara Agustina, Andy Darma, Reza Ranuh, Dadik Raharjo, Toshiro Shirakawa, Masato Fujisawa, and Subijanto Marto Sudarmo. "Clinical manifestation of norovirus infection in children aged less than five years old admitted with acute diarrhea in Surabaya, Indonesia: a cross-sectional study." F1000Research 8 (March 9, 2020): 2130. http://dx.doi.org/10.12688/f1000research.21069.3.
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Background: The objective of this study was to investigate the clinical manifestation of norovirus infection between norovirus genogroup and severity of acute diarrhea in pediatric patients at the Dr. Soetomo Hospital, Surabaya, Indonesia. Methods: This cross-sectional study involved 31 participants aged 1-60 months admitted to the hospital with acute diarrhea from April 2012 to March 2013. Norovirus genogroups (GI and II) were identified from patient stool using reverse transcription polymerase chain reaction (RT-PCR). Severity was measured using the Ruuska and Vesikari scoring system. Results: In total, 94 stool samples were obtained, of which 31 (19%) were norovirus positive. Norovirus GI was found in one sample with mild diarrhea. Norovirus GII was found in 30 samples (96.8%); one sample with mild diarrhea (3.3%), 20 samples with moderate diarrhea (66.7%), and nine samples with severe diarrhea (30%). Conclusion: Norovirus GII was the most prevalent cause of acute diarrhea and 30% of the cases manifested as severe diarrhea.
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König, Katja Marie Kjara, Aminu S. Jahun, Komal Nayak, Lydia N. Drumright, Matthias Zibauer, Ian Goodfellow, and Myra Hosmillo. "Design, development, and validation of a strand-specific RT-qPCR assay for GI and GII human Noroviruses." Wellcome Open Research 6 (September 23, 2021): 245. http://dx.doi.org/10.12688/wellcomeopenres.17078.1.
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Human noroviruses (HuNoV) are the major cause of viral gastroenteritis worldwide. Similar to other positive-sense single-stranded RNA viruses, norovirus RNA replication requires the formation of a negative strand RNA intermediate. Methods for detecting and quantifying the viral positive or negative sense RNA in infected cells and tissues can be used as important tools in dissecting virus replication. In this study, we have established a sensitive and strand-specific Taqman-based quantitative polymerase chain reaction (qPCR) assay for both genogroups GI and GII HuNoV. This assay shows good reproducibility, has a broad dynamic range and is able to detect a diverse range of isolates. We used tagged primers containing a non-viral sequence for the reverse transcription (RT) reaction and targeted this tag in the succeeding qPCR reaction to achieve strand specificity. The specificity of the assay was confirmed by the detection of specific viral RNA strands in the presence of high levels of the opposing strands, in both RT and qPCR reactions. Finally, we further validated the assay in norovirus replicon-bearing cell lines and norovirus-infected human small intestinal organoids, in the presence or absence of small-molecule inhibitors. Overall, we have established a strand-specific qPCR assay that can be used as a reliable method to understand the molecular details of the human norovirus life cycle.
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Fardah Athiyyah, Alpha, Katsumi Shigemura, Koichi Kitagawa, Nazara Agustina, Andy Darma, Reza Ranuh, Dadik Raharjo, Toshiro Shirakawa, Masato Fujisawa, and Subijanto Marto Sudarmo. "Norovirus genogroup correlation with acute diarrhea severity in Indonesian pediatric patients aged 1-60 months: a cross-sectional study." F1000Research 8 (December 20, 2019): 2130. http://dx.doi.org/10.12688/f1000research.21069.1.
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Background: The objective of this study was to investigate the correlation between norovirus genogroup and severity of acute diarrhea in pediatric patients at the Dr. Soetomo Hospital, Surabaya, Indonesia. Methods: This cross-sectional study involved 31 participants aged 1-60 months admitted to the hospital with acute diarrhea from April 2012 to March 2013. Norovirus genogroups (GI and II) were identified from patient stool using reverse transcription polymerase chain reaction (RT-PCR). Severity was measured using the Ruuska and Vesikari scoring system. Results: In total, 91 stool samples were obtained, of which 31 (19%) were norovirus positive. Norovirus GI was found in one sample with mild diarrhea. Norovirus GII was found in 30 samples (96.8%); one sample with mild diarrhea (3.3%), 20 samples with moderate diarrhea (66.7%), and nine samples with severe diarrhea (30%). Conclusion: Norovirus GII was the most prevalent cause of acute diarrhea and 30% of the cases manifested as severe diarrhea.
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Fardah Athiyyah, Alpha, Katsumi Shigemura, Koichi Kitagawa, Nazara Agustina, Andy Darma, Reza Ranuh, Dadik Raharjo, Toshiro Shirakawa, Masato Fujisawa, and Subijanto Marto Sudarmo. "Norovirus genogroup correlation with acute diarrhea severity in Indonesian pediatric patients aged 1-60 months: a cross-sectional study." F1000Research 8 (February 14, 2020): 2130. http://dx.doi.org/10.12688/f1000research.21069.2.
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Background: The objective of this study was to investigate the correlation between norovirus genogroup and severity of acute diarrhea in pediatric patients at the Dr. Soetomo Hospital, Surabaya, Indonesia. Methods: This cross-sectional study involved 31 participants aged 1-60 months admitted to the hospital with acute diarrhea from April 2012 to March 2013. Norovirus genogroups (GI and II) were identified from patient stool using reverse transcription polymerase chain reaction (RT-PCR). Severity was measured using the Ruuska and Vesikari scoring system. Results: In total, 91 stool samples were obtained, of which 31 (19%) were norovirus positive. Norovirus GI was found in one sample with mild diarrhea. Norovirus GII was found in 30 samples (96.8%); one sample with mild diarrhea (3.3%), 20 samples with moderate diarrhea (66.7%), and nine samples with severe diarrhea (30%). Conclusion: Norovirus GII was the most prevalent cause of acute diarrhea and 30% of the cases manifested as severe diarrhea.
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BRUGGINK, L. D., J. M. MOSELEN, and J. A. MARSHALL. "Genotype analysis of noroviruses associated with gastroenteritis outbreaks in childcare centres, Victoria, Australia, 2012–2015." Epidemiology and Infection 145, no. 9 (April 11, 2017): 1933–41. http://dx.doi.org/10.1017/s0950268817000681.
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SUMMARYThe characteristics of norovirus outbreaks in children (0–5 years) in childcare centres in Victoria, Australia (2012–2015) were examined. The three most common open reading frame (ORF) 2 genotypes in childcare centre outbreaks were GII.4 (42%), GII.6 (21%) and GII.3 (14%); the remaining genotypes (GI.2, GI.3, GI.4, GI.8, GI.13, GII.1, GII.2, GII.7 and GII.13) each made up <10%. The GII.4 genotype was the only norovirus genotype seen in all 4 years of the study and was the most common genotype in 2012–2014 but in 2015 the most common genotype was GII.2. The GII.4 genotype was more common in children 0–2 years, whereas GII.2 and GII.7 were more common in children 4–5 years. ORF 1/ORF 2 recombinant forms identified were GII.P4_NewOrleans_2009/GII.4_Sydney_2012, GII.P12/GII.3, GII.Pb (GII.21)/GII.3, GII.Pe/GII.2, GII.Pe/GII.4_Sydney_2012 and GII.Pg/GII.1. The findings indicate that norovirus genotype prevalence patterns in children were influenced by the age of the children and the year in which the analysis was carried out. The majority of norovirus infections (84%) occurred after the first year of life so that vaccination before the age of one would appear to be the most efficacious.
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Shiota, Tomoyuki, Michio Okame, Sayaka Takanashi, Pattara Khamrin, Makiko Takagi, Kenji Satou, Yuichi Masuoka, et al. "Characterization of a Broadly Reactive Monoclonal Antibody against Norovirus Genogroups I and II: Recognition of a Novel Conformational Epitope." Journal of Virology 81, no. 22 (September 12, 2007): 12298–306. http://dx.doi.org/10.1128/jvi.00891-07.
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ABSTRACT Norovirus, which belongs to the family Caliciviridae, is one of the major causes of nonbacterial acute gastroenteritis in the world. The main human noroviruses are of genogroup I (GI) and genogroup II (GII), which were subdivided further into at least 15 and 18 genotypes (GI/1 to GI/15 and GII/1 to GII/18), respectively. The development of immunological diagnosis for norovirus had been hindered by the antigen specificity of the polyclonal antibody. Therefore, several laboratories have produced broadly reactive monoclonal antibodies, which recognize the linear GI and GII cross-reactive epitopes or the conformational GI-specific epitope. In this study, we characterized the novel monoclonal antibody 14-1 (MAb14-1) for further development of the rapid immunochromatography test. Our results demonstrated that MAb14-1 could recognize 15 recombinant virus-like particles (GI/1, 4, 8, and 11 and GII/1 to 7 and 12 to 15) and showed weak affinity to the virus-like particle of GI/3. This recognition range is the broadest of the existing monoclonal antibodies. The epitope for MAb14-1 was identified by fragment, sequence, structural, and mutational analyses. Both terminal antigenic regions (amino acid positions 418 to 426 and 526 to 534) on the C-terminal P1 domain formed the conformational epitope and were in the proximity of the insertion region (positions 427 to 525). These regions contained six amino acids responsible for antigenicity that were conserved among genogroup(s), genus, and Caliciviridae. This epitope mapping explained the broad reactivity and different titers among GI and GII. To our knowledge, we are the first group to identify the GI and GII cross-reactive monoclonal antibody, which recognizes the novel conformational epitope. From these data, MAb14-1 could be used further to develop immunochromatography.
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Lochridge, Vance P., Kathryn L. Jutila, Joel W. Graff, and Michele E. Hardy. "Epitopes in the P2 domain of norovirus VP1 recognized by monoclonal antibodies that block cell interactions." Journal of General Virology 86, no. 10 (October 1, 2005): 2799–806. http://dx.doi.org/10.1099/vir.0.81134-0.
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Noroviruses cause the majority of epidemic outbreaks of acute viral gastroenteritis worldwide. Human norovirus strains do not grow in cell culture, but recent carbohydrate binding, sequence and structural analyses have begun to define functional domains in the norovirus capsid that may be conserved among multiple antigenic types. The purpose of this study was to localize domains and define sequences in the major capsid protein VP1 that are important for cell interactions. Monoclonal antibodies to genogroups GI.1 and GII.2 reference strains Norwalk virus and Snow Mountain virus, respectively, were generated that blocked binding of recombinant virus-like particles to Caco-2 intestinal cells and inhibited haemagglutination. Peptides that mimicked the mAb binding epitopes were selected from a phage-displayed random nonapeptide library. Anti-recombinant Norwalk virus mAb 54.6 and anti-recombinant Snow Mountain virus mAb 61.21 recognized epitopes located in the protruding P2 domain of VP1. The epitope recognized by mAb 61.21 contained amino acids that are completely conserved among norovirus strains across genogroups, including strains isolated from swine, bovine and murine species. This study identifies the first epitope involved in inhibition of norovirus–cell interactions and supports increasing evidence that interactions between noroviruses and host cells rely on structures in the P2 domain of VP1.
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Yang, Siyuan, Min Li, Jingwei Cheng, Gang Wan, Yunao Zhou, Hongyu Jia, Hongshan Wei, et al. "Diagnostic determination of Norovirus infection as one of the major causes of infectious diarrhea in HIV patients using a multiplex polymerase chain reaction assay." International Journal of STD & AIDS 30, no. 6 (February 5, 2019): 550–56. http://dx.doi.org/10.1177/0956462418824912.
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Although infectious diarrhea is one of the most common complications in human immunodeficiency virus (HIV)-infected patients, robust diagnostic methods for determining potential pathogens are still limited in the clinic. Norovirus, a type of calicivirus, has been shown to be the most common cause of gastroenteritis. Here, we used multiplex polymerase chain reaction as a diagnostic tool to verify Norovirus as the diarrhea-related pathogen in HIV-infected patients with unknown etiological information. Stool specimens obtained from 81 HIV-infected patients with diarrhea were analyzed by BioFire FilmArray Gastrointestinal (GI) panel. Among 26 HIV-infected patients with unknown etiological information, we detected Norovirus in 14 stool specimens of these patients with 100% sensitivity and specificity as confirmed by reverse transcription polymerase chain reaction (RT-PCR), and one specimen showed both Norovirus and enterotoxigenic Escherichia coli infection. Among the remaining 55 patients with verified Clostridium difficile infection, nine patients also detected positive for Norovirus infection. In conclusion, using FilmArray GI panel and RT-PCR, we determined that Norovirus infection as one of the main pathogens responsible for diarrhea in HIV patients.
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Nakjarung, Kaewkanya, Ladaporn Bodhidatta, Pimmnapar Neesanant, Paphavee Lertsethtakarn, Orntipa Sethabutr, Ket Vansith, Chhour Y. Meng, Brett E. Swierczewski, and Carl J. Mason. "Molecular Epidemiology and Genetic Diversity of Norovirus in Young Children in Phnom Penh, Cambodia." Journal of Tropical Medicine 2016 (2016): 1–8. http://dx.doi.org/10.1155/2016/2707121.
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This study investigated the genetic diversity of noroviruses identified from a previous surveillance study conducted at the National Pediatric Hospital in Phnom Penh, Cambodia, from 2004 to 2006. In the previous study, 926 stool samples were collected from children aged 3–60 months with acute diarrhea (cases) and without diarrhea (controls) with reported 6.7% of cases and 3.2% of controls being positive for norovirus. The initial norovirus diagnostic assay was performed with real-time reverse transcription-polymerase chain reaction (real-time RT PCR) which also distinguished between genogroups I and II (GI and GII). Norovirus infection was most commonly detected in children aged 12–23 months in both cases and controls. Norovirus Genotyping Tool and phylogenetic analysis of partial sequences of the 3′ end of the RNA-dependent RNA Polymerase (RdRp) and the capsid domain region were employed to assign genotypes of the norovirus strains. GII.4 was the most predominant capsid genotype detected at 39.5% followed by GII.6 at 14.9%. The GII.4 Hunter 2004 variant was the predominant strain detected. Six RdRP/capsid recombinants including GII.P7/GII.6, GII.P7/GII.14, GII.P7/GII.20, GII.P12/GII.13, GII.P17/GII.16, and GII.P21/GII.3 were also identified. This study of norovirus infection in young children in Cambodia suggests genetic diversity of norovirus as reported worldwide.
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Ranuh, Reza Gunadi, Alpha Fardah Athiyyah, Deanty Ayu PA, Andy Darma, Dadik Rahardjo, Toshiro Shirakawa, and Subijanto Marto Sudarmo. "Assessment of the Rapid Immunochromatographic Test as a Diagnostic Tool for Norovirus Related Diarrhea in Children." Folia Medica Indonesiana 55, no. 1 (April 9, 2019): 48. http://dx.doi.org/10.20473/fmi.v55i1.12557.
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In developing countries, Norovirus is the second-leading cause of acute diarrhea, after rotavirus. The approved gold standard method for diagnosis of norovirus infection is RT-PCR. The rapid immunochromatographic test is a novel and expedient method for diagnosing norovirus that is relatively affordable. However, the use of the rapid immunochromatographic test remains controversial because of its accuracy. This study aimed to explore whether the rapid immunochromatographic test could be used for diagnosing norovirus-related diarrhea in children. Rapid immunochromatographic test (QuickNaviTM-Norovirus2) and RT-PCR on stool samples was used to diagnose norovirus. Stool samples were obtained from pediatric patients aged between 1 and 60 months who had diarrhea and were admitted to the pediatric ward at Dr. Soetomo General Hospital Surabaya, between April 2013 and March 2014. Ninety-four subjects provided stool samples that were tested using QuickNaviTM-Noro2 and RT-PCR. Using the test, 64 samples tested positive for norovirus and 30 tested negatives. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the rapid immunochromatographic test were consecutively 90.3%, 42.9%, 43.8%, 90%, and 58.5%. RT-PCR was used to test all samples to assess the accuracy, which showed that one from 31 samples contained the GI strain (1.1%), while 30 samples (32%) contained the GII strain. This study definitively establishes that the rapid immunochromatography test is not sufficiently accurate for use as a screening or diagnostic tool in norovirus-related diarrhea cases in children.
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Ranuh, Reza Gunadi, Alpha Fardah Athiyyah, Deanty Ayu PA, Andy Darma, Dadik Raharjo, Toshiro Shirakawa, and Subijanto Marto Sudarmo. "Assessment of the Rapid Immunochromatographic Test as a Diagnostic Tool for Norovirus Related Diarrhea in Children." Folia Medica Indonesiana 55, no. 1 (January 14, 2021): 48. http://dx.doi.org/10.20473/fmi.v55i1.24377.
Full textAbstract:
In developing countries, Norovirus is the second-leading cause of acute diarrhea, after rotavirus. The approved gold standard method for diagnosis of norovirus infection is RT-PCR. The rapid immunochromatographic test is a novel and expedient method for diagnosing norovirus that is relatively affordable. However, the use of the rapid immunochromatographic test remains controversial because of its accuracy. This study aimed to explore whether the rapid immunochromatographic test could be used for diagnosing norovirus-related diarrhea in children. Rapid immunochromatographic test (QuickNaviTM-Norovirus2) and RT-PCR on stool samples was used to diagnose norovirus. Stool samples were obtained from pediatric patients aged between 1 and 60 months who had diarrhea and were admitted to the pediatric ward at Dr. Soetomo General Hospital Surabaya, between April 2013 and March 2014. Ninety-four subjects provided stool samples that were tested using QuickNaviTM-Noro2 and RT-PCR. Using the test, 64 samples tested positive for norovirus and 30 tested negatives. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the rapid immunochromatographic test were consecutively 90.3%, 42.9%, 43.8%, 90%, and 58.5%. RT-PCR was used to test all samples to assess the accuracy, which showed that one from 31 samples contained the GI strain (1.1%), while 30 samples (32%) contained the GII strain. This study definitively establishes that the rapid immunochromatography test is not sufficiently accurate for use as a screening or diagnostic tool in norovirus-related diarrhea cases in children.
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PUUSTINEN, L., V. BLAZEVIC, L. HUHTI, E. D. SZAKAL, A. HALKOSALO, M. SALMINEN, and T. VESIKARI. "Norovirus genotypes in endemic acute gastroenteritis of infants and children in Finland between 1994 and 2007." Epidemiology and Infection 140, no. 2 (April 14, 2011): 268–75. http://dx.doi.org/10.1017/s0950268811000549.
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SUMMARYNoroviruses are, after rotaviruses, the second most common causative agents of acute gastroenteritis in young children. We studied norovirus genotypes in faecal specimens collected from Finnish children followed-up prospectively in rotavirus vaccine trials. Almost 5000 faecal specimens collected from cases of acute gastroenteritis were examined using reverse transcriptase–PCR. A total of 1172 cases (25% of all acute gastroenteritis) were associated with noroviruses. Of these, 96% were genogroup GII. GII.4 was the most common genotype (46%) throughout the study period but the proportion of this genotype varied in different norovirus epidemic seasons. Additional norovirus genotypes detected were: GII.7 (15%), GII.3 (14%), GII.1 (9%), GII.b (7%), GII.2 (3%), and GI.3 (2%). GII.4 dominated during the following years: 1998–1999 (75%), 2002–2003 (88%) and 2006–2007 (98%) while recombinant genotype GII.b was dominant between 2003 and 2004 (83%). In conclusion, genotypes GII.4 and GIIb have emerged as predominant norovirus genotypes in endemic gastroenteritis affecting young infants and children in Finland.
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Bok, Karin, Gabriel I. Parra, Tanaji Mitra, Eugenio Abente, Charlene K. Shaver, Denali Boon, Ronald Engle, et al. "Chimpanzees as an animal model for human norovirus infection and vaccine development." Proceedings of the National Academy of Sciences 108, no. 1 (December 20, 2010): 325–30. http://dx.doi.org/10.1073/pnas.1014577107.
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Noroviruses are global agents of acute gastroenteritis, but the development of control strategies has been hampered by the absence of a robust animal model. Studies in chimpanzees have played a key role in the characterization of several fastidious hepatitis viruses, and we investigated the feasibility of such studies for the noroviruses. Seronegative chimpanzees inoculated i.v. with the human norovirus strain Norwalk virus (NV) did not show clinical signs of gastroenteritis, but the onset and duration of virus shedding in stool and serum antibody responses were similar to that observed in humans. NV RNA was detected in intestinal and liver biopsies concurrent with the detection of viral shedding in stool, and NV antigen expression was observed in cells of the small intestinal lamina propria. Two infected chimpanzees rechallenged 4, 10, or 24 mo later with NV were resistant to reinfection, and the presence of NV-specific serum antibodies correlated with protection. We evaluated the immunogenicity and efficacy of virus-like particles (VLPs) derived from NV (genogroup I, GI) and MD145 (genogroup II, GII) noroviruses as vaccines. Chimpanzees vaccinated intramuscularly with GI VLPs were protected from NV infection when challenged 2 and 18 mo after vaccination, whereas chimpanzees that received GII VLPs vaccine or a placebo were not. This study establishes the chimpanzee as a viable animal model for the study of norovirus replication and immunity, and shows that NV VLP vaccines could induce protective homologous immunity even after extended periods of time.
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Brown, Julianne R., Sunando Roy, Christopher Ruis, Erika Yara Romero, Divya Shah, Rachel Williams, and Judy Breuer. "Norovirus Whole-Genome Sequencing by SureSelect Target Enrichment: a Robust and Sensitive Method." Journal of Clinical Microbiology 54, no. 10 (August 3, 2016): 2530–37. http://dx.doi.org/10.1128/jcm.01052-16.
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Norovirus full-genome sequencing is challenging due to sequence heterogeneity among genomes. Previous methods have relied on PCR amplification, which is problematic due to primer design, and transcriptome sequencing (RNA-Seq), which nonspecifically sequences all RNA, including host and bacterial RNA, in stool specimens. Target enrichment uses a panel of custom-designed 120-mer RNA baits that are complementary to all publicly available norovirus sequences, with multiple baits targeting each position of the genome, which overcomes the challenge of primer design. Norovirus genomes are enriched from stool RNA extracts to minimize the sequencing of nontarget RNA. SureSelect target enrichment and Illumina sequencing were used to sequence full genomes from 507 norovirus-positive stool samples with reverse transcription–real-time PCR cycle threshold (CT) values of 10 to 43. Sequencing on an Illumina MiSeq system in batches of 48 generated, on average, 81% on-target reads per sample and 100% genome coverage with >12,000-fold read depth. Samples included genotypes GI.1, GI.2, GI.3, GI.6, GI.7, GII.1, GII.2, GII.3, GII.4, GII.5, GII.6, GII.7, GII.13, GII.14, and GII.17. When outliers were accounted for, we generated >80% genome coverage for all positive samples, regardless ofCTvalues. A total of 164 samples were tested in parallel with conventional PCR genotyping of the capsid shell domain; 164/164 samples were successfully sequenced, compared to 158/164 samples that were amplified by PCR. Four of the samples that failed capsid PCR analysis had low titers, which suggests that target enrichment is more sensitive than gel-based PCR. Two samples failed PCR due to primer mismatches; target enrichment uses multiple baits targeting each position, thus accommodating sequence heterogeneity among norovirus genomes.
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Ebenezer, Oluwakemi, Maryam A. Jordaan, Nkululeko Damoyi, and Michael Shapi. "Discovery of Potential Inhibitors for RNA-Dependent RNA Polymerase of Norovirus: Virtual Screening, and Molecular Dynamics." International Journal of Molecular Sciences 22, no. 1 (December 26, 2020): 171. http://dx.doi.org/10.3390/ijms22010171.
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Noroviruses are non-enveloped viruses with a positive-sense single-stranded RNA (ssRNA) genome belonging to the genus Norovirus, from the family Caliciviridae, which are accountable for acute gastroenteritis in humans. The Norovirus genus is subdivided into seven genogroups, i.e., (GI-GVII); among these, the genogroup II and genotype 4 (GII.4) strains caused global outbreaks of human norovirus (HuNov) disease. The viral genome comprises three open reading frames (ORFs). ORF1 encodes the nonstructural polyprotein that is cleaved into six nonstructural proteins, which include 3C-like cysteine protease (3CLpro) and a viral RNA-dependent RNA polymerase. ORF2 and ORF3 encode the proteins VP1 and VP2. The RNA-dependent RNA polymerase (RdRp) from noroviruses is one of the multipurpose enzymes of RNA viruses vital for replicating and transcribing the viral genome, making the virally encoded enzyme one of the critical targets for the development of novel anti-norovirus agents. In the quest for a new antiviral agent that could combat HuNov, high throughput virtual screening (HTVS), combined with e-pharmacophore screening, was applied to screen compounds from the PubChem database. CMX521 molecule was selected as a prototype for a similarity search in the PubChem online database. Molecular dynamics simulations were employed to identify different compounds that may inhibit HuNov. The results predicted that compound CID-57930781 and CID-44396095 formed stable complexes with MNV-RdRp within 50 ns; hence, they may signify as promising human norovirus inhibitors.
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Tohma, Kentaro, Cara J. Lepore, Magaly Martinez, Juan I. Degiuseppe, Pattara Khamrin, Mayuko Saito, Holger Mayta, et al. "Genome-wide analyses of human noroviruses provide insights on evolutionary dynamics and evidence of coexisting viral populations evolving under recombination constraints." PLOS Pathogens 17, no. 7 (July 13, 2021): e1009744. http://dx.doi.org/10.1371/journal.ppat.1009744.
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Norovirus is a major cause of acute gastroenteritis worldwide. Over 30 different genotypes, mostly from genogroup I (GI) and II (GII), have been shown to infect humans. Despite three decades of genome sequencing, our understanding of the role of genomic diversification across continents and time is incomplete. To close the spatiotemporal gap of genomic information of human noroviruses, we conducted a large-scale genome-wide analyses that included the nearly full-length sequencing of 281 archival viruses circulating since the 1970s in over 10 countries from four continents, with a major emphasis on norovirus genotypes that are currently underrepresented in public genome databases. We provided new genome information for 24 distinct genotypes, including the oldest genome information from 12 norovirus genotypes. Analyses of this new genomic information, together with those publicly available, showed that (i) noroviruses evolve at similar rates across genomic regions and genotypes; (ii) emerging viruses evolved from transiently-circulating intermediate viruses; (iii) diversifying selection on the VP1 protein was recorded in genotypes with multiple variants; (iv) non-structural proteins showed a similar branching on their phylogenetic trees; and (v) contrary to the current understanding, there are restrictions on the ability to recombine different genomic regions, which results in co-circulating populations of viruses evolving independently in human communities. This study provides a comprehensive genetic analysis of diverse norovirus genotypes and the role of non-structural proteins on viral diversification, shedding new light on the mechanisms of norovirus evolution and transmission.
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Ji, Zheng, Xiaochang C. Wang, Limei Xu, Chongmiao Zhang, Cheng Rong, Andri Taruna Rachmadi, Mohan Amarasiri, Satoshi Okabe, Naoyuki Funamizu, and Daisuke Sano. "Fecal Source Tracking in A Wastewater Treatment and Reclamation System Using Multiple Waterborne Gastroenteritis Viruses." Pathogens 8, no. 4 (September 30, 2019): 170. http://dx.doi.org/10.3390/pathogens8040170.
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Gastroenteritis viruses in wastewater reclamation systems can pose a major threat to public health. In this study, multiple gastroenteritis viruses were detected from wastewater to estimate the viral contamination sources in a wastewater treatment and reclamation system installed in a suburb of Xi’an city, China. Reverse transcription plus nested or semi-nested PCR, followed by sequencing and phylogenetic analysis, were used for detection and genotyping of noroviruses and rotaviruses. As a result, 91.7% (22/24) of raw sewage samples, 70.8% (17/24) of the wastewater samples treated by anaerobic/anoxic/oxic (A2O) process and 62.5% (15/24) of lake water samples were positive for at least one of target gastroenteritis viruses while all samples collected from membrane bioreactor effluent after free chlorine disinfection were negative. Sequence analyses of the PCR products revealed that epidemiologically minor strains of norovirus GI (GI/14) and GII (GII/13) were frequently detected in the system. Considering virus concentration in the disinfected MBR effluent which is used as the source of lake water is below the detection limit, these results indicate that artificial lake may be contaminated from sources other than the wastewater reclamation system, which may include aerosols, and there is a possible norovirus infection risk by exposure through reclaimed water usage and by onshore winds transporting aerosols containing norovirus.
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Mateo, Roberto, Lisa C. Lindesmith, Shaily J. Garg, Keith Gottlieb, Karen Lin, Sara Said, Juan S. Leon, et al. "Production and Clinical Evaluation of Norwalk GI.1 Virus Lot 001-09NV in Norovirus Vaccine Development." Journal of Infectious Diseases 221, no. 6 (October 20, 2019): 919–26. http://dx.doi.org/10.1093/infdis/jiz540.
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Abstract Background Human noroviruses (HuNoV) are the leading cause of gastroenteritis. No vaccine is currently available to prevent norovirus illness or infection. Safe, infectious challenge strains are needed to assess vaccine efficacy in the controlled human infection model (CHIM). Methods A stock of HuNoV strain Norwalk virus ([NV] GI.1) was prepared. Healthy, genetically susceptible adults were inoculated with NV Lot 001-09NV and monitored for infection, gastroenteritis symptoms, and immune responses. Results Lot 001-09NV induced gastroenteritis in 9 (56%) and infection in 11 (69%) of 16 genetically susceptible subjects. All infected subjects developed strong immune responses to GI.1 with a 30-fold (geometric mean titer) increase in blocking titers (BT50) and a 161-fold increase in GI.1-specific immunoglobulin (Ig)G titers when compared with baseline. GI.1-specific cellular responses in peripheral blood were observed 9 days postchallenge with an average of 3253 IgA and 1227 IgG antibody-secreting cells per million peripheral blood mononuclear cells. Conclusions GI.1 Lot 001-09NV appears to be similar in virulence to previous passages of NV strain 8fIIa. The safety profile, attack rate, and duration of illness make GI.1 Lot 001-09NV a useful challenge strain for future vaccine studies aimed at establishing immune correlates.
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Leshem, Eyal, Leslie Barclay, Mary Wikswo, Everardo Vega, Nicole Gregoricus, Umesh D. Parashar, Jan Vinjé, and Aron J. Hall. "Genotype GI.6 Norovirus, United States, 2010–2012." Emerging Infectious Diseases 19, no. 8 (August 2013): 1317–20. http://dx.doi.org/10.3201/eid1908.130445.
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ZHOU, N., H. ZHANG, X. LIN, P. HOU, S. WANG, Z. TAO, Z. BI, and A. XU. "A waterborne norovirus gastroenteritis outbreak in a school, eastern China." Epidemiology and Infection 144, no. 6 (October 20, 2015): 1212–19. http://dx.doi.org/10.1017/s0950268815002526.
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SUMMARYIn late 2014, a gastroenteritis outbreak occurred in a school in Shandong Province, eastern China. Hundreds of individuals developed the symptoms of diarrhoea and vomiting. Epidemiological investigation showed that food consumption was not linked to this outbreak, and unboiled direct drinking water was identified as the independent risk factor with a relative risk of 1·37 (95% confidence interval 1·03–1·83). Furthermore, examination of common bacterial and viral gastroenteritis pathogens was conducted on different specimens. Norovirus GI.1, GI.2, GI.6, GII.4, GII.6 and GII.13 were detected in clinical specimens and a water sample. GII.4 sequences between clinical specimens and the water sample displayed a close relationship and belonged to GII.4 variant Sydney 2012. These results indicate that direct drinking water contaminated by norovirus was responsible for this gastroenteritis outbreak. This study enriches our knowledge of waterborne norovirus outbreaks in China, and presents valuable prevention and control practices for policy-makers. In future, strengthened surveillance and supervision of direct drinking-water systems is needed.
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Nenonen, Nancy P., Charles Hannoun, Charlotte U. Larsson, and Tomas Bergström. "Marked Genomic Diversity of Norovirus Genogroup I Strains in a Waterborne Outbreak." Applied and Environmental Microbiology 78, no. 6 (January 13, 2012): 1846–52. http://dx.doi.org/10.1128/aem.07350-11.
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ABSTRACTMarked norovirus (NoV) diversity was detected in patient samples from a large community outbreak of gastroenteritis with waterborne epidemiology affecting approximately 2,400 people. NoV was detected in 33 of 50 patient samples examined by group-specific real-time reverse transcription-PCR. NoV genotype I (GI) strains predominated in 31 patients, with mixed GI infections occurring in 5 of these patients. Sequence analysis of RNA-dependent polymerase-N/S capsid-coding regions (∼900 nucleotides in length) confirmed the dominance of the GI strains (n= 36). Strains of NoV GI.4 (n= 21) and GI.7 (n= 9) were identified, but six strains required full capsid amino acid analyses (530 to 550 amino acids) based on control sequencing of cloned amplicons before the virus genotype could be determined. Three strains were assigned to a new NoV GI genotype, proposed as GI.9, based on capsid amino acid analyses showing 26% dissimilarity from the established genotypes GI.1 to GI.8. Three other strains grouped in a sub-branch of GI.3 with 13 to 15% amino acid dissimilarity to GI.3 GenBank reference strains. Phylogenetic analysis (2.1 kb) of 10 representative strains confirmed these genotype clusters. Strains of NoV GII.4 (n= 1), NoV GII.6 (n= 2), sapovirus GII.2 (n= 1), rotavirus (n= 3), adenovirus (n= 1), andCampylobacterspp. (n= 2) were detected as single infections or as mixtures with NoV GI. Marked NoV GI diversity detected in patients was consistent with epidemiologic evidence of waterborne NoV infections, suggesting human fecal contamination of the water supply. Recognition of NoV diversity in a cluster of patients provided a useful warning marker of waterborne contamination in the Lilla Edet outbreak.
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Crawford, Sue E., Nadim Ajami, Tracy Dewese Parker, Noritoshi Kitamoto, Katsuro Natori, Naokazu Takeda, Tomoyuki Tanaka, Baijun Kou, Robert L. Atmar, and Mary K. Estes. "Mapping Broadly Reactive Norovirus Genogroup I and II Monoclonal Antibodies." Clinical and Vaccine Immunology 22, no. 2 (November 26, 2014): 168–77. http://dx.doi.org/10.1128/cvi.00520-14.
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ABSTRACTNoroviruses are responsible for most acute nonbacterial epidemic outbreaks of gastroenteritis worldwide. To develop cross-reactive monoclonal antibodies (MAbs) for rapid identification of genogroup I and II (GI and GII) noroviruses (NoVs) in field specimens, mice were immunized with baculovirus-expressed recombinant virus-like particles (VLPs) corresponding to NoVs. Nine MAbs against the capsid protein were identified that detected both GI and GII NoV VLPs. These MAbs were tested in competition enzyme-linked immunosorbent assays (ELISAs) to identify common epitope reactivities to GI and GII VLPs. Patterns of competitive reactivity placed these MAbs into two epitope groups (groups 1 and 2). Epitopes for MAbs NV23 and NS22 (group 1) and MAb F120 (group 2) were mapped to a continuous region in the C-terminal P1 subdomain of the capsid protein. This domain is within regions previously defined to contain cross-reactive epitopes in GI and GII viruses, suggesting that common epitopes are clustered within the P1 domain of the capsid protein. Further characterization in an accompanying paper (B. Kou et al., Clin Vaccine Immunol 22:160–167, 2015,http://dx.doi.org/10.1128/CVI.00519-14) revealed that MAb NV23 (epitope group 1) is able to detect GI and GII viruses in stool. Inclusion of the GI and GII cross-reactive MAb NV23 in antigen detection assays may facilitate the identification of GI and GII human noroviruses in stool samples as causative agents of outbreaks and sporadic cases of gastroenteritis worldwide.