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      Effect of thermal and chemical treatments used for SARS-COV-2 inactivation in the measurement of saliva analytes

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          Abstract

          The present study aims to assess the effects of thermal and chemical inactivating procedures, that can be used for SARS-CoV-2 inactivation, on different salivary analytes. SDS–Polyacrylamide Gel Electrophoresis (SDS-PAGE) protein profile and a panel of 25 specific biomarkers of oxidative status, stress, metabolism and tissue damage were evaluated in samples subjected to different treatments: thermal (65 °C or 92 °C) and chemical with detergents [sodium dodecyl sulphate (SDS), Triton X-100 or NP-40]. Salivary SDS-PAGE profile was most affected by heating at 92 °C, with three and two protein bands decreasing and increasing their expression levels, respectively. This treatment also affected the results of several enzymes, with some of them being also affected by heating at 65 °C and incubation with SDS. The use of Triton X-100 or NP-40 resulted in increased values of cortisol, triglycerides and glucose, not affecting the other tested biomarkers. The present results will help researchers and clinicians to select the best protocols to work in safe conditions with saliva, taking into account the target analyte planned to be measured.

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          A pneumonia outbreak associated with a new coronavirus of probable bat origin

          Since the outbreak of severe acute respiratory syndrome (SARS) 18 years ago, a large number of SARS-related coronaviruses (SARSr-CoVs) have been discovered in their natural reservoir host, bats 1–4 . Previous studies have shown that some bat SARSr-CoVs have the potential to infect humans 5–7 . Here we report the identification and characterization of a new coronavirus (2019-nCoV), which caused an epidemic of acute respiratory syndrome in humans in Wuhan, China. The epidemic, which started on 12 December 2019, had caused 2,794 laboratory-confirmed infections including 80 deaths by 26 January 2020. Full-length genome sequences were obtained from five patients at an early stage of the outbreak. The sequences are almost identical and share 79.6% sequence identity to SARS-CoV. Furthermore, we show that 2019-nCoV is 96% identical at the whole-genome level to a bat coronavirus. Pairwise protein sequence analysis of seven conserved non-structural proteins domains show that this virus belongs to the species of SARSr-CoV. In addition, 2019-nCoV virus isolated from the bronchoalveolar lavage fluid of a critically ill patient could be neutralized by sera from several patients. Notably, we confirmed that 2019-nCoV uses the same cell entry receptor—angiotensin converting enzyme II (ACE2)—as SARS-CoV.
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            Inactivation of the coronavirus that induces severe acute respiratory syndrome, SARS-CoV

            Severe acute respiratory syndrome (SARS) is a life-threatening disease caused by a novel coronavirus termed SARS-CoV. Due to the severity of this disease, the World Health Organization (WHO) recommends that manipulation of active viral cultures of SARS-CoV be performed in containment laboratories at biosafety level 3 (BSL3). The virus was inactivated by ultraviolet light (UV) at 254 nm, heat treatment of 65 °C or greater, alkaline (pH > 12) or acidic (pH < 3) conditions, formalin and glutaraldehyde treatments. We describe the kinetics of these efficient viral inactivation methods, which will allow research with SARS-CoV containing materials, that are rendered non-infectious, to be conducted at reduced safety levels.
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              Saliva is a reliable tool to detect SARS-CoV-2

              Highlights • Saliva is a reliable tool to detect SARS-Cov-2 by RT-rPCR analysis. • Saliva may provide information about the clinical evolution of the disease. • Saliva could represent a valid instrument in COVID-19 diagnosis. • Patients should be checked for salivary viral load at hospital discharge.
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                Author and article information

                Contributors
                ecsl@uevora.pt
                jjceron@um.es
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                8 June 2022
                8 June 2022
                2022
                : 12
                : 9434
                Affiliations
                [1 ]GRID grid.8389.a, ISNI 0000 0000 9310 6111, MED-Mediterranean Institute for Agriculture, Environment and Development & CHANGE-Global Change and Sustainability Institute, , University of Évora, ; Ap. 94, 7006-554 Évora, Portugal
                [2 ]GRID grid.10586.3a, ISNI 0000 0001 2287 8496, Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Veterinary School, Regional Campus of International Excellence Mare Nostrum, , University of Murcia, ; Campus de Espinardo s/n, Espinardo, 30100 Murcia, Spain
                [3 ]GRID grid.8389.a, ISNI 0000 0000 9310 6111, Department of Medical and Health Sciences, School of Health and Human Development, , University of Évora, ; Évora, Portugal
                [4 ]GRID grid.10586.3a, ISNI 0000 0001 2287 8496, Interdisciplinary Laboratory of Clinical Analysis, Interlab-UMU, Regional Campus of International Excellence Mare Nostrum, , University of Murcia, ; Espinardo, 30100 Murcia, Spain
                [5 ]GRID grid.411101.4, ISNI 0000 0004 1765 5898, Faculty of Medicine and Odontology, , Hospital Morales Meseguer, Clínica Odontológica, ; Marqués del los Vélez s/n, 30008 Murcia, Spain
                Article
                13491
                10.1038/s41598-022-13491-9
                9174913
                35676391
                3a430bcd-a0ec-4cc8-b409-5e38eee82d6b
                © The Author(s) 2022

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 20 November 2021
                : 17 May 2022
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100001871, Fundação para a Ciência e a Tecnologia;
                Award ID: UIDB/05183/2020
                Categories
                Article
                Custom metadata
                © The Author(s) 2022

                Uncategorized
                diagnostic markers,predictive markers,biochemical assays,proteomic analysis
                Uncategorized
                diagnostic markers, predictive markers, biochemical assays, proteomic analysis

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