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      Investigation of SARS-CoV-2 Detection Method Applicability and Virus Occurrence in Food and Food Packaging

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          SUMMARY

          Research background

          While it is clear that SARS CoV-2 coronavirus is the primary respiratory virus, there are no entirely clarified ways of transmission. Foodborne transmission has remained an unexplained path. Therefore, the goals of this paper are to examine and present an assessment of the most appropriate of the four selected kits for RNA extraction for the testing and detection of SARS-CoV-2 on food packaging surfaces, food surfaces, and in food. This will enable to indicate the possibility of infection through contact or direct food consumption.

          Experimental approach

          Finding the best technique is vital as RNA extraction is one of the essential elements in detecting SARS-CoV-2. This was achieved through an experiment with four commercial kits following the original manufacturers’ protocols, and with a modification of the original protocols that included the use of ethanol and isopropanol. The selected kit was used for RNA extraction from the swabs of packaging surfaces, food surface, and ready-to-eat food samples. The coronavirus was then identified using real-time reverse transcription-polymerase chain reaction (RT-PCR) assays to determine whether the SARS-CoV-2 virus or viral particles are present in the food chain with the overall purpose of demonstrating the possibility that food can contribute as a vehicle for the transmission of the virus.

          Results and conclusions

          The findings of this investigation made the most effective extraction kit and protocol stand out. The results of the applicability of the kit indicated a significant share of positive samples of viral SARS-CoV-2 virus particles on surfaces from the environment where infected persons with 'silent' COVID-19 infection, with mild symptoms or no symptoms, were present. However, according to the findings of the second part of the study, the virus was not detected on the examined samples of food packaging surfaces, food surfaces, and food.

          Novelty and scientific contribution

          The presented results distinguished one of the most suitable protocols for isolating RNA from environmental surface samples. The main contribution of the study is in the presentation of the results, that is, the examination of samples that are primarily related to the food chain, food packaging, food surfaces, and ready-to-eat food. The results of this study could also be helpful for further determination of the potential of food as a vector for the transmission of coronaviruses.

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          Most cited references32

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          Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR

          Background The ongoing outbreak of the recently emerged novel coronavirus (2019-nCoV) poses a challenge for public health laboratories as virus isolates are unavailable while there is growing evidence that the outbreak is more widespread than initially thought, and international spread through travellers does already occur. Aim We aimed to develop and deploy robust diagnostic methodology for use in public health laboratory settings without having virus material available. Methods Here we present a validated diagnostic workflow for 2019-nCoV, its design relying on close genetic relatedness of 2019-nCoV with SARS coronavirus, making use of synthetic nucleic acid technology. Results The workflow reliably detects 2019-nCoV, and further discriminates 2019-nCoV from SARS-CoV. Through coordination between academic and public laboratories, we confirmed assay exclusivity based on 297 original clinical specimens containing a full spectrum of human respiratory viruses. Control material is made available through European Virus Archive – Global (EVAg), a European Union infrastructure project. Conclusion The present study demonstrates the enormous response capacity achieved through coordination of academic and public laboratories in national and European research networks.
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            Air, Surface Environmental, and Personal Protective Equipment Contamination by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) From a Symptomatic Patient

            This study documents results of SARS-CoV-2 polymerase chain reaction (PCR) testing of environmental surfaces and personal protective equipment surrounding 3 COVID-19 patients in isolation rooms in a Singapore hospital.
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              Stability of SARS-CoV-2 in different environmental conditions

              We previously reported the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in different clinical samples. 1 This virus can be detected on different surfaces in a contaminated site. 2 Here, we report the stability of SARS-CoV-2 in different environmental conditions. We first measured the stability of SARS-CoV-2 at different temperatures. SARS-CoV-2 in virus transport medium (final concentration ∼6·8 log unit of 50% tissue culture infectious dose [TCID50] per mL) was incubated for up to 14 days and then tested for its infectivity (appendix p 1). The virus is highly stable at 4°C, but sensitive to heat. At 4°C, there was only around a 0·7 log-unit reduction of infectious titre on day 14. With the incubation temperature increased to 70°C, the time for virus inactivation was reduced to 5 mins. We further investigated the stability of this virus on different surfaces. Briefly, a 5 μL droplet of virus culture (∼7·8 log unit of TCID50 per mL) was pipetted on a surface (appendix p 1; ∼cm2 per piece) and left at room temperature (22°C) with a relative humidity of around 65%. The inoculated objects retrieved at desired time-points were immediately soaked with 200 μL of virus transport medium for 30 mins to elute the virus. Therefore, this recovery of virus does not necessarily reflect the potential to pick up the virus from casual contact. No infectious virus could be recovered from printing and tissue papers after a 3-hour incubation, whereas no infectious virus could be detected from treated wood and cloth on day 2. By contrast, SARS-CoV-2 was more stable on smooth surfaces. No infectious virus could be detected from treated smooth surfaces on day 4 (glass and banknote) or day 7 (stainless steel and plastic). Strikingly, a detectable level of infectious virus could still be present on the outer layer of a surgical mask on day 7 (∼0·1% of the original inoculum). Interestingly, a biphasic decay of infectious SARS-CoV-2 could be found in samples recovered from these smooth surfaces (appendix pp 2–7). 39 representative non-infectious samples tested positive by RT-PCR 3 (data not shown), showing that non-infectious viruses could still be recovered by the eluents. We also tested the virucidal effects of disinfectants by adding 15 μL of SARS-CoV-2 culture (∼7·8 log unit of TCID50 per mL) to 135 μL of various disinfectants at working concentration (appendix p 1). With the exception of a 5-min incubation with hand soap, no infectious virus could be detected after a 5-min incubation at room temperature (22°C). Additionally, we also found that SARS-CoV-2 is extremely stable in a wide range of pH values at room temperature (pH 3–10; appendix p 1). Overall, SARS-CoV-2 can be highly stable in a favourable environment, 4 but it is also susceptible to standard disinfection methods.
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                Author and article information

                Journal
                Food Technol Biotechnol
                Food Technol Biotechnol
                FTB
                Food Technology and Biotechnology
                University of Zagreb Faculty of Food Technology and Biotechnology
                1330-9862
                1334-2606
                June 2023
                June 2023
                : 61
                : 2
                : 250-258
                Affiliations
                [1 ]Andrija Štampar Teaching Institute of Public Health , Mirogojska cesta 16, 10000 Zagreb, Croatia
                [2 ]deptFaculty of Food Technology and Biotechnology , University of Zagreb , Pierottijeva ulica 6, 10000 Zagreb, Croatia
                [3 ]deptFaculty of Medicine , University of Rijeka , Braće Branchetta 20, 51 000 Rijeka, Croatia
                Author notes
                [* ]Corresponding author: Phone: +38514696246, E-mail: zdenko.mlinar@ 123456stampar.hr

                AUTHORS' CONTRIBUTION

                Z. Mlinar developed and conceptualized the idea and methodology of the study, conducted the experiment, analysed data, interpreted statistical analysis, and wrote the main draft of the manuscript. D. Kostelac participated in experiment preparation and writing of the original draft. J. Frece and I. Kovaček contributed to the experiment preparation, manuscript editing, and critical revision. A. Klobučar and V. Tešić contributed to the experiment preparation, field sampling in the quarantine, and critical revision of the manuscript. V. Prahin conducted the field sampling of all final study samples and participated in the implementation of tests for the presence of viruses. Z. Mlinar prepared the figures. All authors reviewed the manuscript.

                Author information
                https://orcid.org/0000-0003-1428-5468
                https://orcid.org/0000-0002-1120-2965
                https://orcid.org/0000-0002-9462-5073
                https://orcid.org/0000-0003-3464-6830
                https://orcid.org/0000-0003-2499-9931
                https://orcid.org/0000-0003-4742-7856
                https://orcid.org/0000-0003-1022-4377
                Article
                FTB-61-250
                10.17113/ftb.61.02.23.8018
                10339731
                f7280043-04c9-4392-9322-cfa16e60d02a
                Copyright @ 2023

                This is an open-access article distributed under the terms of the Creative Commons Attribution (CC BY) 4.0 License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 30 November 2022
                : 13 June 2023
                Categories
                Preliminary Communications

                sars-cov-2,food,rt-pcr,coronavirus,foodborne transmission,rna extraction

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