Tag Archives: reverse transcription pcr

Research – Norovirus – Surfaces – Handlers – DIsinfectants

Ingenta ConnectEurofins

Human infective noroviruses (NoVs) are a worldwide leading cause of foodborne illness and are frequently spread via infected food handlers preparing and manipulating food products such as deli sandwiches. The objective of the current study was to determine the efficiencies whereby NoV could be transferred between surfaces associated with the preparation of manually prepared foods such as deli sandwiches. Nonfood surfaces included gloves and stainless steel discs, and boiled ham, lettuce, and a sandwich bun were the ingredients of the deli sandwich. Both NoV GII.4 and the murine NoV 1 (MNV-1, a cultivable human NoV surrogate) were included in the presented study. Transfer of NoV GII.4 and MNV-1 between surfaces was performed by pressing an inoculated donor surface against an acceptor surface. To evaluate the effect of subsequent contact, donor surfaces were pressed a second time to an identical acceptor surface. Subsequently, NoV GII.4 and MNV-1 were detected using real-time reverse transcription PCR assays and plaque assays, respectively. Transfer of both viruses from gloves to stainless steel was inefficient, and virus transfer from food products to stainless steel occurred with more variability for NoV GII.4 than for MNV-1. Virus transfer from the stainless steel discs to the gloves was substantially more efficient than from the gloves to the stainless steel. NoV GII.4 and MNV-1 transfer from food products to the gloves occurred with varying efficiencies, although this variation was more evident for NoV GII.4. The MNV-1 inoculum was significantly less efficiently transferred to the acceptor surface at the second contact, which was not the case for NoV GII.4. The obtained transfer efficiency data may provide insights into the transfer of NoV during preparation of foods and can be included in risk assessment models describing the transmission of NoVs in this context.

Ingenta Connect

Human noroviruses (HuNoVs) are the most common cause of acute viral gastroenteritis worldwide and are a leading cause of foodborne disease. Their environmental persistence and purported resistance to disinfection undoubtedly contribute to their success as foodborne disease agents. The purpose of this study was to compare the efficacy of three commonly used disinfectant active ingredients against representative HuNoV strains and cultivable surrogates. Ethanol (50, 70, and 90%), sodium hypochlorite (5, 75, 250, 500, and 1,000 ppm), and a quaternary ammonium compound blend (at 0.1×, 1.0×, and 10× concentrations) were evaluated against two norovirus (NoV) genogroup II strains (GII.2 and GII.4) and two surrogates (feline calicivirus [FCV] and murine norovirus [MNV-1]). Virucidal suspension assays (30-s exposure) were conducted in accordance with ASTM International standard E-1052. Virus inactivation was quantified using reverse transcription quantitative PCR targeting the ORFI-ORFII junction (HuNoV), the RNA polymerase region (MNV-1), or the ORFI region (FCV); infectivity assays were also performed for MNV-1 and FCV. The two HuNoV strains and FCV were relatively resistant to ethanol (<0.5 log inactivation) irrespective of concentration, but MNV-1 was much more susceptible (log inactivation, ∼2.0 log at higher ethanol concentrations). Both HuNoV strains were more resistant to hypochlorite than were either of the animal surrogates, with the human strains requiring ≥500 ppm of hypochlorite to achieve statistically significant reduction (≥3.0 log) in virus concentration. All four viruses were resistant to inactivation (<0.5-log reduction) using the quaternary ammonium compound formulation at all concentrations tested. This study is novel in that it clearly demonstrates the relative ineffectiveness of common active disinfectant ingredients against HuNoV and highlights the fact that the cultivable surrogates do not always mimic HuNoV strains.