Extraction of human noroviruses from leafy greens and fresh herbs using magnetic silica beads

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creativework.keywords - en
RNA extraction
PCR inhibitors
Process control
Capsid integrity
dc.contributor.author
Raymond, Philippe
Paul, Sylvianne
Perron, André
Deschênes, Louise
Hara, Kenji
dc.date.accessioned
2023-05-15T15:17:39Z
dc.date.available
2023-05-15T15:17:39Z
dc.date.issued
2021-05-11
dc.description.abstract - en
Consumption of leafy greens and to a lesser extent fresh herbs has been associated with several foodborne outbreaks including human norovirus (HuNoV). However, the extraction and detection of viruses from these matrices present multiple challenges such as low recovery yields and relatively high PCR inhibition. A new magnetic silica bead based (MSB) extraction protocol was developed and used to recover norovirus from leafy greens and fresh herbs. The performance results were compared to the ISO 15216–1:2017 standard. The HuNoV GII.4 and GI.5 recovery yields from spiked lettuce using the MSB extraction protocol range from 33 to 82%. There was a good correlation between murine norovirus (MNV) and HuNoV recovery yields from fresh herbs and leafy greens. No reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR) inhibition was detected from leafy green extracts using the MSB methodology. The selected commercial RT-qPCR detection kit had a major impact on RT-qPCR inhibition levels detected in the ISO 15216–1:2017 RNA extracts. RNase treatment was used to estimate genome recovery from HuNoV with intact capsids. This treatment resulted in similar HuNoV and MNV recovery yields. Between 2019 and 2020, the MSB protocol was used to conduct a survey of HuNoV in domestic and imported leafy greens and fresh herbs sold at retail in Canada. All of the 280 samples tested were negative. Overall, the use of MSB was shown to be an efficient approach to recover HuNoV from leafy greens and certain types of fresh herbs and to conduct surveys.
dc.identifier.citation
Raymond, P., Paul, S., Perron, A., Deschênes, L., & Hara, K. (2021). Extraction of human noroviruses from leafy greens and fresh herbs using magnetic silica beads. Food Microbiology, 99. https://doi.org/10.1016/j.fm.2021.103827
dc.identifier.doi
https://doi.org/10.1016/j.fm.2021.103827
dc.identifier.issn
1095-9998
dc.identifier.uri
https://open-science.canada.ca/handle/123456789/284
dc.language.iso
en
dc.publisher
Elsevier
dc.rights - en
None
dc.rights - fr
Aucune
dc.rights.openaccesslevel - en
Green
dc.rights.openaccesslevel - fr
Vert
dc.rights.uri - en
No License
dc.rights.uri - fr
Aucune Licence
dc.subject - en
Agriculture
dc.subject - fr
Agriculture
dc.subject.en - en
Agriculture
dc.subject.fr - fr
Agriculture
dc.title - en
Extraction of human noroviruses from leafy greens and fresh herbs using magnetic silica beads
dc.type - en
Article
dc.type - fr
Article
local.article.journaltitle
Food Microbiology
local.article.journalvolume
99
local.peerreview - en
Yes
local.peerreview - fr
Oui
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