High-Pressure Inactivation of Human Norovirus Virus-Like Particles Provides Evidence that the Capsid of Human Norovirus Is Highly Pressure Resistant

Lou, Fangfei; Huang, Pengwei; Neetoo, Hudaa; Gurtler, Joshua B.; Niemira, Brendan A.; Chen, Chien-Jen; Jiang, Xi; Lǐ, Jiànróng

doi:10.1128/aem.00532-12

Cited by 44 publications

(41 citation statements)

References 38 publications

(93 reference statements)

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“…As shown in Fig. 3, and consistent with data described previously (20), two protein bands, with molecular masses of ϳ60 and ϳ50 kDa, corresponding to full-length VP1 and the cleaved form of VP1 (cVP1), respectively, were detected in VLPs. Exposure to stainless steel surfaces resulted in virtually no reduction in the HuNoV VP1 band intensity, regardless of time (Fig.…”

Section: Resultssupporting

confidence: 74%

Destruction of the Capsid and Genome of GII.4 Human Norovirus Occurs during Exposure to Metal Alloys Containing Copper

Manuel

Moore

Jaykus

2015

Appl Environ Microbiol

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Human norovirus (HuNoV) represents a significant public health burden worldwide and can be environmentally transmitted. Copper surfaces have been shown to inactivate the cultivable surrogate murine norovirus, but no such data exist for HuNoV. The purpose of this study was to characterize the destruction of GII.4 HuNoV and virus-like particles (VLPs) during exposure to copper alloy surfaces. Fecal suspensions positive for a GII.4 HuNoV outbreak strain or GII.4 VLPs were exposed to copper alloys or stainless steel for 0 to 240 min and recovered by elution. HuNoV genome integrity was assessed by reverse transcriptionquantitative PCR (RT-qPCR) (without RNase treatment), and capsid integrity was assessed by RT-qPCR (with RNase treatment), transmission electron microscopy (TEM), SDS-PAGE/Western blot analysis, and a histo-blood group antigen (HBGA) binding assay. Exposure of fecal suspensions to pure copper for 60 min reduced the GII.4 HuNoV RNA copy number by ϳ3 log 10 units when analyzed by RT-qPCR without RNase treatment and by 4 log 10 units when a prior RNase treatment was used. The rate of reduction of the HuNoV RNA copy number was approximately proportional to the percentage of copper in each alloy. Exposure of GII.4 HuNoV VLPs to pure-copper surfaces resulted in noticeable aggregation and destruction within 240 min, an 80% reduction in the VP1 major capsid protein band intensity in 15 min, and a near-complete loss of HBGA receptor binding within 8 min.In all experiments, HuNoV remained stable on stainless steel. These results suggest that copper surfaces destroy HuNoV and may be useful in preventing environmental transmission of the virus in at-risk settings.H uman norovirus (HuNoV) is the leading cause of viral gastroenteritis worldwide (1, 2). In the United States alone, the virus is responsible for 19 million to 21 million illnesses annually, contributing to nearly $500 million in hospital-associated costs each year (3, 4). The illness is characterized by nausea, vomiting, and diarrhea and typically lasts 24 to 48 h. While the disease is usually self-limiting, in individuals belonging to sensitive populations, symptoms can become life threatening if rehydration therapy is ignored.HuNoV transmission occurs via the fecal-oral route, usually through ingestion of contaminated food or water, or by direct contact with an infected individual. Environmental transmission also occurs, and episodes of vomiting or diarrhea can contaminate surfaces with infectious virus particles that may persist for weeks (5). Environmental persistence of HuNoV is enhanced by its resistance to a variety of commonly used sanitizers and disinfectants, for example, alcohol-based hand sanitizers and hypochlorite at regulated concentrations (6, 7). These unique traits of HuNoV, combined with the low infectious dose and the fact that infected individuals shed a large amount of virus in both fecal material and vomitus, contribute to the high number of outbreaks observed annually in environments with close quarters such as cruise ships, restau...

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Section: Resultssupporting

confidence: 74%

Destruction of the Capsid and Genome of GII.4 Human Norovirus Occurs during Exposure to Metal Alloys Containing Copper

Manuel

Moore

Jaykus

2015

Appl Environ Microbiol

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“…It is generally recognized that surrogates and hNoV survive in a variety of raw, processed, and/or stored foods, including berries, herbs, and molluscan shellfish (Baert et al, 2009;Verhaelen et al, 2012;Richards, 2012;Richards et al, 2012;Poschetto et al, 2007;Butot et al, 2008;Li et al, 2014;Sánchez et al, 2011;Lou et al, 2012). Studies have also shown that the food matrix and environmental factors have a significant impact on virus survival (Le Guyader and Atmar, 2008;Sobsey and Meschke, 2003).…”

Section: Citationmentioning

confidence: 97%

A systematic review of human norovirus survival reveals a greater persistence of human norovirus RT-qPCR signals compared to those of cultivable surrogate viruses

Knight

Haines

Stals

et al. 2016

International Journal of Food Microbiology

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“…However, a major concern is that the results of real-time RT-PCR do not accurately reflect the level of viral infectivity. We previously found that the mechanism of viral inactivation by HPP is the disruption of the viral capsid and not the degradation of genomic RNA, and this capsid disruption would directly affect the receptor binding ability (14,33). Thus, intact virus particles that possess receptor binding activity should be able to bind to PGM-MBs and subsequently be detected by real-time RT-PCR.…”

Section: Discussionmentioning

confidence: 99%

A Gnotobiotic Pig Model for Determining Human Norovirus Inactivation by High-Pressure Processing

Lou

et al. 2015

Appl Environ Microbiol

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e Human norovirus (NoV) is responsible for over 90% of outbreaks of acute nonbacterial gastroenteritis worldwide and accounts for 60% of cases of foodborne illness in the United States. Currently, the infectivity of human NoVs is poorly understood due to the lack of a cell culture system. In this study, we determined the survival of a human NoV genogroup II, genotype 4 (GII.4) strain in seeded oyster homogenates after high-pressure processing (HPP) using a novel receptor binding assay and a gnotobiotic pig model. Pressure conditions of 350 MPa at 0°C for 2 min led to a 3.7-log 10 reduction in the number of viral RNA copies in oysters, as measured by the porcine gastric mucin-conjugated magnetic bead (PGM-MB) binding assay and real-time RT-PCR, whereas pressure conditions of 350 MPa at 35°C for 2 min achieved only a 1-log 10 reduction in the number of RNA copies. Newborn gnotobiotic piglets orally fed oyster homogenate treated at 350 MPa and 0°C for 2 min did not have viral RNA shedding in feces, histologic lesions, or viral replication in the small intestine. In contrast, gnotobiotic piglets fed oysters treated at 350 MPa and 35°C for 2 min had high levels of viral shedding in feces and exhibited significant histologic lesions and viral replication in the small intestine. Collectively, these data demonstrate that (i) human NoV survival estimated by an in vitro PGM-MB virus binding assay is consistent with the infectivity determined by an in vivo gnotobiotic piglet model and (ii) HPP is capable of inactivating a human NoV GII.4 strain at commercially acceptable pressure levels. Human norovirus (NoV), a member of the Caliciviridae family, is responsible for over 90% of the outbreaks of acute nonbacterial gastroenteritis worldwide and accounts for more than 60% of the cases of foodborne illness in the United States (1, 2). It is estimated that 48 million individuals, or about 17% of the U.S. population, are sickened each year, leading to approximately 128,000 hospitalizations and 3,000 fatalities in the United States (2). Human NoV is transmitted primarily through the fecal-oral route either by direct person-to-person contact or by fecally contaminated food or water. Human NoV is highly contagious and stable, and only a few virus particles are thought to be sufficient to cause an infection (3, 4). Outbreaks frequently occur in restaurants, hotels, day care centers, schools, nursing homes, cruise ships, swimming pools, hospitals, and military installations. Despite the significant economic impact and high morbidity caused by human NoV, no vaccines or antiviral drugs with activity against this virus are currently available (5, 6). This is due in large part to the lack of a cell culture system and a small-animal model for human NoV (6, 7). As a consequence, the survival of human NoV is poorly understood.A major high-risk food for human NoV contamination is seafood, particularly bivalves, such as oysters, mussels, and clams (8-11). These animals are filter feeders and can readily bioaccumulate human NoV in their tissue...

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High-Pressure Inactivation of Human Norovirus Virus-Like Particles Provides Evidence that the Capsid of Human Norovirus Is Highly Pressure Resistant

Cited by 44 publications

References 38 publications

Destruction of the Capsid and Genome of GII.4 Human Norovirus Occurs during Exposure to Metal Alloys Containing Copper

Destruction of the Capsid and Genome of GII.4 Human Norovirus Occurs during Exposure to Metal Alloys Containing Copper

A systematic review of human norovirus survival reveals a greater persistence of human norovirus RT-qPCR signals compared to those of cultivable surrogate viruses

A Gnotobiotic Pig Model for Determining Human Norovirus Inactivation by High-Pressure Processing

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