Binding of Virus-Like Particles of Norwalk Virus to Romaine Lettuce Veins

Gandhi, Kamal M.; Mandrell, Robert E.; Tian, Peng

doi:10.1128/aem.01566-10

Cited by 47 publications

(35 citation statements)

References 26 publications

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“…Although it is known that NoVs can attach to lettuce leaves (14,49), the underlying factors that facilitate this interaction are not clear. In the current study, we demonstrated that a NoV-lettuce interaction is mediated by various carbohydrate moieties that are present in the plant cell walls.…”

Section: Discussionmentioning

confidence: 99%

Binding of Human GII.4 Norovirus Virus-Like Particles to Carbohydrates of Romaine Lettuce Leaf Cell Wall Materials

Esseili

Wang

Saif

2012

Appl Environ Microbiol

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Norovirus (NoV) genogroup II genotype 4 (GII.4) strains are the dominant cause of the majority of food-borne outbreaks, including those that involve leafy greens, such as lettuce. Since human NoVs use carbohydrates of histo-blood group antigens as receptors/coreceptors, we examined the role of carbohydrates in the attachment of NoV to lettuce leaves by using virus-like particles (VLPs) of a human NoV/GII.4 strain. Immunofluorescence analysis showed that the VLPs attached to the leaf surface, especially to cut edges, stomata, and along minor veins. Binding was quantified using enzyme-linked immunosorbent assay (ELISA) performed on cell wall materials (CWM) from innermost younger leaves and outermost lamina of older leaves. The binding to CWM of older leaves was significantly (P < 0.05) higher (1.5-to 2-fold) than that to CWM of younger leaves. Disrupting the carbohydrates of CWM or porcine gastric mucin (PGM) (a carbohydrate control) using 100 mM sodium periodate (NaIO 4 ) significantly decreased the binding an average of 17% in younger leaves, 43% in older leaves, and 92% for PGM. In addition, lectins recognizing GalNAc, GlcNAc, and sialic acid at 100 g/ml significantly decreased the binding an average of 41%, 33%, and 20% on CWM of older leaves but had no effect on younger leaves. Lectins recognizing ␣-D-Gal, ␣-D-Man/␣-D-Glc, and ␣-L-Fuc showed significant inhibition on CWM of older leaves as well as that of younger leaves. All lectins, except for the lectin recognizing ␣-D-Gal, significantly inhibited NoV VLP binding to PGM. Collectively, our results indicate that NoV VLPs bind to lettuce CWM by utilizing multiple carbohydrate moieties. This binding may enhance virus persistence on the leaf surface and prevent effective decontamination.

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Section: Discussionmentioning

confidence: 99%

Binding of Human GII.4 Norovirus Virus-Like Particles to Carbohydrates of Romaine Lettuce Leaf Cell Wall Materials

Esseili

Wang

Saif

2012

Appl Environ Microbiol

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“…Other researchers have developed ELISAs for VLP detection using variable antigen ranges. Esseili and coworkers reported an assay for GII.4 VLP that can detect 0.05-2.50 μg (50 μL aliquots of 1, 5, 10, 20, and 50 μg/mL VLPs) [31], while Ghandi et al [32] created an ELISA assay for GI.1 VLP that used 100 μg (100 μL aliquots of 1.0 μg/mL total protein) [32]. The GI.1 VLPbased ELISA developed at the Baylor College of Medicine -the basis for the assays developed in this study -used 0.63, 1.25, 2.5, and 5 μg in the initial antigen binding step [28].…”

Section: Resultsmentioning

confidence: 99%

“…Current approaches for detecting norovirus in clinical and environmental samples utilize a combination of electron microscopy techniques [20][21][22][23][24], molecular detection assays [7,10,17,[25][26][27] and immunological methods [25,[28][29][30][31][32][33]. Each approach strives for maximizing sensitivity and specificity while minimizing the time for detection [34].…”

Section: Introductionmentioning

confidence: 99%

“…ELISAs can analyze a myriad of pathological samples using multiple screening modes and reporter molecules, giving the assay broad versatility, but their need for high viral loads [5,27,30] limits the assays' application mostly in clinical settings. Despite their relatively lower sensitivity when compared to PCR-based methods [5,20], ELISA-based assays have been used to detect norovirus in human stool samples [5,25,[36][37][38][39][40][41][42][43], human sera [29], and food samples [26,31,32,44,45] Using norovirus GI.1 (Norwalk) virus-like particles (VLPs) as a model viral system, the objective of this study was to develop and evaluate several ELISA-based assays for rapid detection of varying concentrations of GI.1 VLPs (0.037-3.7 μg/mL). HuNoV VLPs are replication-incompetent, macromolecular protein assemblies with capsid structures and antigenic properties resembling those of innate norovirus particles [46][47][48].…”

Section: Introductionmentioning

confidence: 99%

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Design and Evaluation of Three Immuno-based Assays for Rapid Detection of Human Norovirus Virus-like Particles

Broglie¹,

Moore²,

Jaykus³

et al. 2014

J Anal Bioanal Tech

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This study designed and evaluated three versatile immuno-based assays for the rapid detection of GI.1 norovirus virus-like particles at low (0-3.0 μg/mL) levels: 1) enzymatic absorbance-based ELISAs, 2) a fluorescentbased immunoassay, and 3) a "signal-down" capture ELISA. Variables including controllable variations in assay format (indirect or sandwich), assay time, binding sequence, and reporter molecule (fluorophore or enzyme) were thoroughly investigated and optimized in all three assays. Selectivity tests in the three-hour absorbance-based ELISA using VLPs representing two GI and two GII strains indicated the assays were selective to GI strains over GII strains. The three-hour enzymatic absorbance-based assay turned out to be a robust and rapid method capable of detecting GI.I VLPs in the range of 0.037 to 0.555 μg/mL, and the three-hour fluorescent immunoassay was capable of detecting VLPs in a high concentration range of 0.5-2.0 μg/mL under optimized conditions. The "signal-down" capture ELISA was conceptually demonstrated for the detection of VLPs at concentrations in excess of 1.0 μg/ mL, but did not appear to be suitable for quantifying VLPs under its current conditions. The methods reported here provide proof-of-concept that various ELISA-type approaches could be further developed to provide robust norovirus detection assays having various detection ranges, limits, and linearity.

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“…Furthermore, other biological factors, such as leaf leachates (12,69), may impact the survival of both viruses and bacteria on leaves. However, since virus-like particles of HuNoV were previously reported to adhere to certain locations on the plant phyllosphere (14,70,71) where native bacteria are reported to adhere, such as the leaf veins and stomata (72), it is possible that the HuNoVs contaminating plant leaves occupy niches similar to those that native bacteria occupy and therefore directly interact with those bacteria. A recent study by our group has shown an enhanced persistence of SaV on postharvest lettuce leaves that were infected by a plant-pathogenic bacterium preharvest (73), suggesting that plant bacteria may have a direct influence on the survival of viruses on the leaves.…”

Section: Discussionmentioning

confidence: 99%

Abiotic Stress and Phyllosphere Bacteria Influence the Survival of Human Norovirus and Its Surrogates on Preharvest Leafy Greens

Esseili

Gao

Tegtmeier

et al. 2016

Appl Environ Microbiol

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Foodborne outbreaks of human noroviruses (HuNoVs) are frequently associated with leafy greens. Because there is no effective method to eliminate HuNoV from postharvest leafy greens, understanding virus survival under preharvest conditions is crucial. The objective of this study was to evaluate the survival of HuNoV and its surrogate viruses, murine norovirus (MNV), porcine sapovirus (SaV), and Tulane virus (TV), on preharvest lettuce and spinach that were subjected to abiotic stress (physical damage, heat, or flood). We also examined the bacteria culturable from the phyllosphere in response to abiotic stress and in relation to viral persistence. Mature plants were subjected to stressors 2 days prior to inoculation of the viruses on leaves. We quantified the viral RNA, determined the infectivity of the surrogates, and performed bacterial counts on postinoculation days (PIDs) 0, 1, 7, and 14. For both plant types, time exerted significant effects on HuNoV, MNV, SaV, and TV RNA titers, with greater effects being seen for the surrogates. Infectious surrogate viruses were undetectable on PID 14. Only physical damage on PID 14 significantly enhanced HuNoV RNA persistence on lettuce, while the three stressors differentially enhanced the persistence of MNV and TV RNA. Bacterial counts were significantly affected by time and plant type but not by the stressors. However, bacterial counts correlated significantly with HuNoV RNA titers on spinach and with the presence of surrogate viruses on both plant types under various conditions. In conclusion, abiotic stressors and phyllosphere bacterial density may differentially influence the survival of HuNoV and its surrogates on lettuce and spinach, emphasizing the need for the use of preventive measures at the preharvest stage.H uman noroviruses (HuNoVs) are the leading cause of acute gastroenteritis and foodborne illnesses in the United States, causing 19 million to 21 million cases, 570 to 800 deaths, and $777 million in health care-associated costs annually (1). These singlestranded RNA viruses belong to the Caliciviridae family and are 28 to 35 nm in diameter and nonenveloped. Although these viruses cause self-limiting gastroenteritis, more serious infections can develop in high-risk groups, such as the elderly and immunocompromised populations (2). The virus is mainly transmitted to humans via the fecal-oral route either by contact with an infected person or fomites or by ingestion of contaminated food and water (2). Leafy greens are frequently associated with HuNoV outbreaks (1) and are globally recognized to be a high priority in terms of the microbial safety of fresh produce (3). Contamination of leafy greens with HuNoVs can occur at any stage along the farm-to-fork chain (3) through a number of sources, including fecally contaminated water used for irrigation, improperly treated sewage sludge used for fertilization, and food harvesters or food handlers who use improper hygiene practices (4, 5). Because HuNoVs have a low infectious dose (ϳ18 to 1,000 viral particles) (6) ...

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Binding of Virus-Like Particles of Norwalk Virus to Romaine Lettuce Veins

Cited by 47 publications

References 26 publications

Binding of Human GII.4 Norovirus Virus-Like Particles to Carbohydrates of Romaine Lettuce Leaf Cell Wall Materials

Binding of Human GII.4 Norovirus Virus-Like Particles to Carbohydrates of Romaine Lettuce Leaf Cell Wall Materials

Design and Evaluation of Three Immuno-based Assays for Rapid Detection of Human Norovirus Virus-like Particles

Abiotic Stress and Phyllosphere Bacteria Influence the Survival of Human Norovirus and Its Surrogates on Preharvest Leafy Greens

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