Norovirus Extraction from Frozen Raspberries Using Magnetic Silica Beads

Philippe Raymond,Louise Deschênes,Sylvianne Paul,André Perron

doi:10.1007/s12560-021-09466-0

Philippe Raymond, Louise Deschênes + Show 2 more

Open Access

https://doi.org/10.1007/s12560-021-09466-0

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Abstract

Human noroviruses (HuNoV) are among the main causes of acute gastroenteritis worldwide. Frozen raspberries have been linked to several HuNoV food-related outbreaks. However, the extraction of HuNoV RNA from frozen raspberries remains challenging. Recovery yields are low, and real-time quantitative reverse transcriptase PCR (RT-qPCR) inhibitors limit the sensitivity of the detection methodologies. A new approach using fine magnetic silica beads was developed for the extraction of HuNoV spiked on frozen raspberries. Relatively low recovery yields were observed with both the magnetic silica bead and the reference ISO 15216-1:2017 methods. High RT-qPCR inhibition was observed with the ISO 15216-1:2017 recommended amplification kit but could be reduced by using an alternative kit. Reducing RT-qPCR inhibition is important to limit the number of inconclusive HuNoV assays thus increasing the capacity to assess the HuNoV prevalence in frozen raspberries.

Highlights

Human Norovirus (HuNoV) is one of the leading causes of food-related illnesses in developed countries
The performance of the magnetic silica bead (MSB) method was evaluated by assessing its recovery yields in comparison to the ISO 15216-1:2017 method using individually quick frozen (IQF) frozen raspberries spiked with HuNoV GII.4, GI.5, or Murine norovirus-1 (MNV) (Table 2)
The virus elution and concentration methods for norovirus using the MSB had a turnaround time of approximately 7 h, including 1 h required for the robotic RNA extraction

Summary

Introduction

Human Norovirus (HuNoV) is one of the leading causes of food-related illnesses in developed countries. HuNoV represents 58% of all reported foodborne outbreaks of known etiology in the United States (Vinje 2015). About 300–400 outbreaks of HuNoV are reported to the National Enteric Surveillance Program of the Public Health Agency of Canada each year (Government of Canada 2018). Forty-six berry outbreaks associated to HuNoV contamination with 15,827 cases were reported globally between 1983 and 2018 (Bozkurt et al 2020). In the European Union, contaminated frozen red fruits were shown to represent an important cause of HuNoV outbreaks (Boqvist et al 2018). Frozen raspberries were implicated as food vehicles in 33 of the 40 reported outbreak events associated

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