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      Rapid electrochemical detection of coronavirus SARS-CoV-2

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          Abstract

          Coronavirus disease 2019 (COVID-19) is a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Diagnosis of COVID-19 depends on quantitative reverse transcription PCR (qRT-PCR), which is time-consuming and requires expensive instrumentation. Here, we report an ultrasensitive electrochemical biosensor based on isothermal rolling circle amplification (RCA) for rapid detection of SARS-CoV-2. The assay involves the hybridization of the RCA amplicons with probes that were functionalized with redox active labels that are detectable by an electrochemical biosensor. The one-step sandwich hybridization assay could detect as low as 1 copy/μL of N and S genes, in less than 2 h. Sensor evaluation with 106 clinical samples, including 41 SARS-CoV-2 positive and 9 samples positive for other respiratory viruses, gave a 100% concordance result with qRT-PCR, with complete correlation between the biosensor current signals and quantitation cycle (Cq) values. In summary, this biosensor could be used as an on-site, real-time diagnostic test for COVID-19.

          Abstract

          Currently the most common method of COVID-19 diagnosis is by qRT-PCR which is slow and requires expensive instrumentation. Here the authors report an electrochemical biosensor based on isothermal rolling circle amplification for rapid detection of SARS-CoV-2 in clinical samples.

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          Most cited references41

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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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            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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              Detection of SARS-CoV-2 in Different Types of Clinical Specimens

              This study describes results of PCR and viral RNA testing for SARS-CoV-2 in bronchoalveolar fluid, sputum, feces, blood, and urine specimens from patients with COVID-19 infection in China to identify possible means of non-respiratory transmission.
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                Author and article information

                Contributors
                benchaporn.ler@mahidol.ac.th
                Journal
                Nat Commun
                Nat Commun
                Nature Communications
                Nature Publishing Group UK (London )
                2041-1723
                5 February 2021
                5 February 2021
                2021
                : 12
                : 802
                Affiliations
                [1 ]GRID grid.10223.32, ISNI 0000 0004 1937 0490, Biosensors Laboratory, Department of Biomedical Engineering, Faculty of Engineering, , Mahidol University, ; Nakhon Pathom, Thailand
                [2 ]GRID grid.7922.e, ISNI 0000 0001 0244 7875, Center of Excellence in Clinical Virology, Faculty of Medicine, , Chulalongkorn University, ; Bangkok, Thailand
                [3 ]GRID grid.412151.2, ISNI 0000 0000 8921 9789, Department of Biotechnology, School of Bioresources and Technology, , King Mongkut’s University of Technology Thonburi, ; Bangkok, Thailand
                [4 ]GRID grid.412151.2, ISNI 0000 0000 8921 9789, Pilot Plant Development and Training Institute (PDTI), , King Mongkut’s University of Technology Thonburi, ; Bangkok, Thailand
                [5 ]GRID grid.1024.7, ISNI 0000000089150953, School of Chemistry and Physics, , Queensland University of Technology (QUT), ; Brisbane, QLD Australia
                [6 ]GRID grid.444449.d, ISNI 0000 0004 0627 9137, Faculty of Applied Sciences, , AIMST University, ; Bedong, Kedah Malaysia
                [7 ]GRID grid.444449.d, ISNI 0000 0004 0627 9137, Centre of Excellence for Omics-Driven Computational Biodiscovery (COMBio), , AIMST University, ; Bedong, Kedah Malaysia
                Author information
                http://orcid.org/0000-0001-9294-5180
                http://orcid.org/0000-0003-0086-8371
                Article
                21121
                10.1038/s41467-021-21121-7
                7864991
                33547323
                16221e73-94ab-471a-a414-8d739be5ce12
                © The Author(s) 2021

                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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 25 September 2020
                : 11 January 2021
                Funding
                Funded by: National Research Council of Thailand, the Research Chair Grant from the National Science and Technology Development Agency (P–15–5004)
                Categories
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                © The Author(s) 2021

                Uncategorized
                oligonucleotide probes,sensors
                Uncategorized
                oligonucleotide probes, sensors

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