Impact of Capsid and Genomic Integrity Tests on Norovirus Extraction Recovery Rates

Human norovirus (HuNoV) is the leading pathogen responsible for food-borne illnesses. However, both infectious and non-infectious HuNoV can be detected by RT-qPCR. This study evaluated the efficiency of different capsid integrity treatments coupled with RT-qPCR or a long-range viral RNA (long RT-qPC...

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Bibliographic Details
Published in:Foods 2023-02, Vol.12 (4), p.826
Main Authors: Raymond, Philippe, Paul, Sylvianne, Guy, Rebecca A
Format: Article
Language:English
Subjects:
capsid integrity
Causes of
chelating agent
Deactivation
Extraction (Chemistry)
Food contamination & poisoning
Food science
Foodborne diseases
Genetic aspects
Genomes
Heat
Heat recovery
Heat treatment
Identification and classification
Integrity
intercalating dye
Lettuce
long RT-qPCR
Methods
Norovirus
Physiological aspects
Polyethylene glycol
Polymerase chain reaction
Ribonuclease
Ribonucleic acid
Risk reduction
RNA
RNase
Structure
Viral proteins
Viruses
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Summary:Human norovirus (HuNoV) is the leading pathogen responsible for food-borne illnesses. However, both infectious and non-infectious HuNoV can be detected by RT-qPCR. This study evaluated the efficiency of different capsid integrity treatments coupled with RT-qPCR or a long-range viral RNA (long RT-qPCR) detection to reduce the recovery rates of heat inactivated noroviruses and fragmented RNA. The three capsid treatments evaluated (RNase, the intercalating agent PMAxx and PtCl ) reduced the recovery of heat inactivated HuNoV and murine norovirus (MNV) spiked on lettuce, when combined with the ISO 15216-1:2017 extraction protocols. However, PtCl also reduced non-heat-treated noroviruses recovery as estimated by RT-qPCR. The PMAxx and RNase treatments had a similar effect on MNV only. The most efficient approaches, the RNase and PMAxx treatments, reduced the heat-inactivated HuNoV recovery rates estimated using RT-qPCR by 2 and >3 log, respectively. The long RT-qPCR detection approach also reduced the recovery rates of heat inactivated HuNoV and MNV by 1.0 and 0.5 log, respectively. Since the long-range viral RNA amplification could be applied to verify or confirm RT-qPCR results, it also provides some advantages by reducing the risk of false positive HuNoV results.
ISSN:2304-8158
2304-8158
DOI:10.3390/foods12040826