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

Author:

Raymond Philippe1ORCID,Paul Sylvianne1ORCID,Guy Rebecca2ORCID

Affiliation:

1. St-Hyacinthe Laboratory—Food Virology, Canadian Food Inspection Agency (CFIA), St-Hyacinthe, QC J2S 8E3, Canada

2. National Microbiology Laboratory, Division of Enteric Diseases, Public Health Agency of Canada (PHAC), Guelph, ON N1G 3W4, Canada

Abstract

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 PtCl4) 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, PtCl4 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.

Funder

Canadian Food Inspection Agency Research & Partnership Strategy

Publisher

MDPI AG

Subject

Plant Science,Health Professions (miscellaneous),Health (social science),Microbiology,Food Science

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