Droplet Digital PCR for Precise Quantification of Human Norovirus in Shellfish Associated with Gastroenteritis Illness
Norovirus is the predominant cause of viral gastroenteritis globally with foodborne outbreaks commonly reported. Filter-feeding bivalve molluscan shellfish can become contaminated with norovirus when grown in waters impacted by inadequately treated effluent wastewater, overflows, or other human feca...
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Elsevier
2024-11-01
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| Series: | Journal of Food Protection |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0362028X24001479 |
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| author | Daniel Rexin Laetitia Kaas Jérémie Langlet Dawn Croucher Joanne Hewitt |
| author_facet | Daniel Rexin Laetitia Kaas Jérémie Langlet Dawn Croucher Joanne Hewitt |
| author_sort | Daniel Rexin |
| collection | DOAJ |
| description | Norovirus is the predominant cause of viral gastroenteritis globally with foodborne outbreaks commonly reported. Filter-feeding bivalve molluscan shellfish can become contaminated with norovirus when grown in waters impacted by inadequately treated effluent wastewater, overflows, or other human fecal sources. Contaminated shellfish pose a significant risk to consumers, because combined with a low norovirus infectious dose, oysters and mussels are often eaten raw or lightly cooked resulting in no or minimal virus inactivation, respectively. In addition, shellfish contamination has significant economic impacts on the seafood industry. To improve risk assessments, reverse transcription (RT)-digital droplet PCR (ddPCR) was used to determine the precise norovirus concentrations in 20 shellfish samples, all positive for norovirus genogroup I and/or II (GI or GII) by RT-quantitative PCR (qPCR), and associated with reported norovirus illness in New Zealand. Using RT-ddPCR, total norovirus GI and/or GII concentrations in shellfish ranged between 44 and 4,630 genome copies (GC)/g digestive tissue. Importantly, 40% (8/20) of shellfish samples contained a total norovirus concentration less than 200 GC/g digestive tissue. In parallel, RNase treatment was applied, prior to viral extraction to remove free viral RNA, which subsequently led to average reductions in norovirus GC/g concentration of 37.1% and 19.4% for GI and GII, respectively. These RT-ddPCR data provide valuable evidence for risk assessment of contaminated shellfish and evaluation of safety guidelines and highlight issues associated with setting a safe threshold of norovirus in shellfish. |
| format | Article |
| id | doaj-art-cdd108fa5e1e4ccbbd4f1f323d438566 |
| institution | OA Journals |
| issn | 0362-028X |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Elsevier |
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| series | Journal of Food Protection |
| spelling | doaj-art-cdd108fa5e1e4ccbbd4f1f323d4385662025-08-20T01:47:26ZengElsevierJournal of Food Protection0362-028X2024-11-01871110036310.1016/j.jfp.2024.100363Droplet Digital PCR for Precise Quantification of Human Norovirus in Shellfish Associated with Gastroenteritis IllnessDaniel Rexin0Laetitia Kaas1Jérémie Langlet2Dawn Croucher3Joanne Hewitt4Institute of Environmental Science and Research Limited (ESR), Enteric, Environmental and Food Virology Laboratory, Porirua, New ZealandInstitute of Environmental Science and Research Limited (ESR), Enteric, Environmental and Food Virology Laboratory, Porirua, New ZealandInstitute of Environmental Science and Research Limited (ESR), Enteric, Environmental and Food Virology Laboratory, Porirua, New ZealandInstitute of Environmental Science and Research Limited (ESR), Enteric, Environmental and Food Virology Laboratory, Porirua, New ZealandCorresponding author at: Enteric, Environmental and Food Virology Laboratory, Institute of Environmental Science and Research Limited (ESR), Kenepuru Science Centre, PO BOX 50-348, Porirua 5240, New Zealand.; Institute of Environmental Science and Research Limited (ESR), Enteric, Environmental and Food Virology Laboratory, Porirua, New ZealandNorovirus is the predominant cause of viral gastroenteritis globally with foodborne outbreaks commonly reported. Filter-feeding bivalve molluscan shellfish can become contaminated with norovirus when grown in waters impacted by inadequately treated effluent wastewater, overflows, or other human fecal sources. Contaminated shellfish pose a significant risk to consumers, because combined with a low norovirus infectious dose, oysters and mussels are often eaten raw or lightly cooked resulting in no or minimal virus inactivation, respectively. In addition, shellfish contamination has significant economic impacts on the seafood industry. To improve risk assessments, reverse transcription (RT)-digital droplet PCR (ddPCR) was used to determine the precise norovirus concentrations in 20 shellfish samples, all positive for norovirus genogroup I and/or II (GI or GII) by RT-quantitative PCR (qPCR), and associated with reported norovirus illness in New Zealand. Using RT-ddPCR, total norovirus GI and/or GII concentrations in shellfish ranged between 44 and 4,630 genome copies (GC)/g digestive tissue. Importantly, 40% (8/20) of shellfish samples contained a total norovirus concentration less than 200 GC/g digestive tissue. In parallel, RNase treatment was applied, prior to viral extraction to remove free viral RNA, which subsequently led to average reductions in norovirus GC/g concentration of 37.1% and 19.4% for GI and GII, respectively. These RT-ddPCR data provide valuable evidence for risk assessment of contaminated shellfish and evaluation of safety guidelines and highlight issues associated with setting a safe threshold of norovirus in shellfish.http://www.sciencedirect.com/science/article/pii/S0362028X24001479Food safetyNorovirusOutbreakPublic healthShellfish |
| spellingShingle | Daniel Rexin Laetitia Kaas Jérémie Langlet Dawn Croucher Joanne Hewitt Droplet Digital PCR for Precise Quantification of Human Norovirus in Shellfish Associated with Gastroenteritis Illness Journal of Food Protection Food safety Norovirus Outbreak Public health Shellfish |
| title | Droplet Digital PCR for Precise Quantification of Human Norovirus in Shellfish Associated with Gastroenteritis Illness |
| title_full | Droplet Digital PCR for Precise Quantification of Human Norovirus in Shellfish Associated with Gastroenteritis Illness |
| title_fullStr | Droplet Digital PCR for Precise Quantification of Human Norovirus in Shellfish Associated with Gastroenteritis Illness |
| title_full_unstemmed | Droplet Digital PCR for Precise Quantification of Human Norovirus in Shellfish Associated with Gastroenteritis Illness |
| title_short | Droplet Digital PCR for Precise Quantification of Human Norovirus in Shellfish Associated with Gastroenteritis Illness |
| title_sort | droplet digital pcr for precise quantification of human norovirus in shellfish associated with gastroenteritis illness |
| topic | Food safety Norovirus Outbreak Public health Shellfish |
| url | http://www.sciencedirect.com/science/article/pii/S0362028X24001479 |
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