Simple and Rapid Detection of Human Norovirus from Produce Using SYBR Green I-based Real-time RT-PCR

Radin, Dragoslava; D’Souza, Doris H.

doi:10.1007/s12560-011-9066-5

Cited by 13 publications

(14 citation statements)

References 38 publications

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“…This sensitivity was higher than the one described by Scipioni and co-workers ( 29 ), who obtained linearity in NoV amplification in human stool samples in the range of 5.8×10 6 to 5.8×10 11 copies. Similar results were also obtained by other authors ( 30 ), who developed a SYBR Green-based real-time RT-PCR for NoV detection in lettuce, cherry tomatoes, and green onions. Recently, Oshiki and co-workers developed a microfluidic nested PCR amplification method for NoV detection in oysters and found that the assay could detect 10 2 -10 5 copies/g digestive tissue ( 31 ).…”

Section: Discussionsupporting

confidence: 89%

Absence of norovirus contamination in shellfish harvested and commercialized in the Northeast coast of Brazil

Guarines

Mendes

Cordeiro

et al. 2020

Braz J Med Biol Res

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Norovirus (NoV) is the main cause of gastroenteritis outbreaks worldwide. Although NoV spreads mainly from person to person, it is estimated that a large proportion of NoV outbreaks are caused by foodborne transmission. Bivalve mollusks are one of the most important foods involved in NoV transmission to humans. Little is known about NoV prevalence in shellfish harvested and commercialized in Brazil. The aim of this study was to map, for the first time, the distribution of NoV contamination in oysters and mussels harvested and commercialized in the coast of Pernambuco state, northeast Brazil. A total of 380 mollusks (260 oysters and 120 mussels) were collected between February and August 2017 either directly from harvesting areas or obtained from beach vendors at 17 sites in Pernambuco. Samples were processed and tested for NoV contamination using a SYBR Green real-time PCR assay. All samples were negative for NoV GI or GII contamination, suggesting a low risk of NoV contamination from this food source during the study period. Additional surveys in different areas of the Brazilian coast are warranted to monitor the risk of NoV infection upon seafood consumption.

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Section: Discussionsupporting

confidence: 89%

Absence of norovirus contamination in shellfish harvested and commercialized in the Northeast coast of Brazil

Guarines

Mendes

Cordeiro

et al. 2020

Braz J Med Biol Res

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“…Some researchers consider that lettuce have the highest adsorption capacity and the most favorable conditions for viral persistence because of the size and the wrinkled texture of its leaves, compared to other smooth, flat surfaces; while others suggest that capacity of lettuce leaf to protect the virus against desiccation is less than green onions or coarse surfaces like ham [17]. In our studies of norovirus GI and GII detection from deli meat, tomato and lettuce similar detection limits of 10 RT-PCRU/25 g were obtained while for green onion limit was 1 RT-PCRU/25g [18,19]. To conclude, viruses can survive on the surface of fresh produce once they have been harvested and remain infectious for several days or for up to 5 weeks during commercial and household storage [20].…”

Section: Transmission and Associated Foodssupporting

confidence: 59%

Noroviruses in food chain production

Radin¹

2016

Hrana i ishrana

Self Cite

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Viruses, predominantly noroviruses, are progressively identified as the leading cause of foodborne diseases worldwide. In the Europe Union they have been determined as a main cause of foodborne outbreaks in 2014, while in the United States viruses are responsible for 58% (5.5 million) of foodborne illnesses annually. Noroviruses, the etiological agent of foodborne gastroenteritis belong to the Caliciviridae family, genus Norovirus which is genetically classified into 6 established geno-groups (GI to GVI). Human noroviruses are icosahedral nonenveloped RNA viruses, approximately 27 to 38 nm in diameter, and contain single-stranded positive-sense RNA genomes size from 7.4 to 8.3 kb and 3 open reading frames. Viruses are transmitted by contaminated food or water, through person-to-person contact, and by cross-contamination from surfaces. Foods at risk includes those which need extensive handling, mostly ready-to-eat foods that do not undergo further processing, and those exposed to environmental contamination, such as seafood and fresh produce. Contamination with human norovirus can happen throughout the entire food production chain: (i) at the primary production (due to fecal polluted water in the harvest area/ irrigation/ water used to dilute pesticide, organic-based fertilizers and potential root uptake of pathogens following internalization); (ii) during harvest (contact with human feces or fecal soiled materials, cross-contamination with equipment and food handlers poor hygiene practice); (iii) at the postharvest stage by inappropriate practices during handling, processing, preparation, storage, distribution (by infected food handlers, spraying with contaminated water, cross-contamination during washing process) and (iv) at the point of sale/consumption (by infected food handlers, cross-contamination with polluted working instruments or surfaces previously contaminated by infected food handlers or food items). Strategies to reduce human noroviruses in the food chain should, beside good hygiene practice, engage interventions to prevent their adsorption to food/contact surfaces and inactivation on/within the products using successful processing techniques.

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“…A total of 122 fecal specimens were collected and tested. The highest dilution of real-time RT-PCR to generate a positive cycle threshold (C t ) signal was one realtime RT-PCR unit (RT-qPCRU) [13]. C t values versus log 10 viral genomic copies linear standard curves were generated from a continuous 10-fold dilution (See Figure S3, Additional le 5).…”

Section: Source Of Hunovs Clinical Samples (Including Suspected Hunovs)mentioning

confidence: 99%

“…HuNoVs are highly-infectious, and infected patients can shed high titers of virus particles with strong resistance to the environmental factors [10,11]. As cell culturing in vitro is still immature to replicate HuNoVs effectively [12], the detection of the virus mainly depends on the molecular approaches such as RT-PCR and RT-qPCR [13,14]. These molecular approaches require specialized equipment, unique reagents, and signi cant time for sample preparation and assay execution, all of which make them impractical for either clinical use or on-site assays of eld samples requiring quick results.…”

Section: Introductionmentioning

confidence: 99%

Broad-range and Effective Detection of Human Noroviruses by Colloidal Gold Immunochromatographic Assay Based on the Shell Domain of the Major Capsid Protei

et al. 2020

Preprint

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Background: Human noroviruses (HuNoVs) are a major cause of nonbacterial gastroenteritis in all ages worldwide. As the replication of HuNoVs in vitro is immature, the detection of HuNoVs is depended on molecular assays such as RT-PCR and RT-qPCR. However, these molecular-based techniques require special equipment, unique reagents, experienced operators to perform, and extended time to get results. In addition, the genotypes of HuNoVs are broad. Therefore, a method for rapidly, broad-range, and effective approach for HuNoVs would be more applicable for screening infected people when outbreaks occur.Results: In this study, a colloidal-gold-based immunochromatographic assay (ICA) was developed for highly effective detection of HuNoVs in clinical samples. Monoclonal antibodies (McAbs) against the shell (S) domain in the major capsid protein of HuNoV were used in the ICA. The limitations of detection for HuNoVs in clinical samples were 1.2×106 genomic copies per gram (gc/g) and 4.4×105 gc/g for genogroup I and II (GI and GII) HuNoVs, respectively. A total of 122 samples were tested for HuNoVs by ICA and compared against that by RT-qPCR. The relative sensitivity, specificity and agreement of the ICA was 84.2 % (95% CI: 83.6-84.8 %), 100.0 % (95 % CI: 98.5-100.0 %) and 87.7 % (95% CI: 85.6-89.8 %), respectively. No cross-reaction with other common enteric viruses or bacteria was observed. The ICA could detect a broad range of genotypes, including GI.1, GI.3, GI.4, GI.6, GI.14, GII.2, GII.3, GII.4, GII.6, GII.13, and GII.17 HuNoVs.Conclusions: Our results demonstrated that ICA targeting the S domain protein is a promising candidate for effectively improve identifying different genotypes of HuNoVs in clinical samples with high sensitivity and specificity.

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Simple and Rapid Detection of Human Norovirus from Produce Using SYBR Green I-based Real-time RT-PCR

Cited by 13 publications

References 38 publications

Absence of norovirus contamination in shellfish harvested and commercialized in the Northeast coast of Brazil

Absence of norovirus contamination in shellfish harvested and commercialized in the Northeast coast of Brazil

Noroviruses in food chain production

Broad-range and Effective Detection of Human Noroviruses by Colloidal Gold Immunochromatographic Assay Based on the Shell Domain of the Major Capsid Protei

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