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      Recent advances and perspectives of nucleic acid detection for coronavirus

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          Abstract

          The recent pneumonia outbreak caused by a novel coronavirus (SARS-CoV-2) in Chinais posing a great threat to global public health. Therefore, rapid and a accurate identification of pathogenic viruses plays a vital role in selecting appropriate treatments, saving people’s lives and preventing epidemics. It is important to establish a quick standard diagnostic test for the detection of the infectious disease (COVID-19) to prevent subsequent secondary spread. Polymerase chain reaction (PCR) is regarded as a gold standard test for the molecular diagnosis of viral and bacterial infections with high sensitivity and specificity. Isothermal nucleic acid amplification is considered to be a highly promising candidate method due to its fundamental advantage in quick procedure time at constant temperature without thermocycler operation. A variety of improved or new approaches also have been developed. This review summarizes the currently available detection methods for coronavirus nucleic acid. It is anticipated that this will assist researchers and clinicians in developing better techniques for timely and effective detection of coronavirus infection.

          Graphical abstract

          The recent pneumonia outbreak caused by a novel coronavirus (SARS-CoV-2) in China, poses a great threat to global public health. Therefore, rapid and accurate identification of pathogenic viruses plays a vital role in selecting appropriate treatments, saving people’s lives and preventing epidemics. This review summarizes the currently available detection methods for coronavirus nucleic acid. It is anticipated that this will assist researchers and clinicians in developing better techniques for timely and effective detection of coronavirus infection.

          Highlights

          • This review summarizes the currently available detection methods for coronavirus nucleic acid.

          • It will assist researchers in developing better techniques for timely and effective detection of coronavirus infection.

          • It will help the establishment of SARS-CoV-2 RNA detection method which is useful for the early diagnosis of COVID-19.

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          Most cited references32

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          Is Open Access

          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 loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation.

            The loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method that uses only one type of enzyme. One of the characteristics of the LAMP method is its ability to synthesize extremely large amount of DNA. Accordingly, a large amount of by-product, pyrophosphate ion, is produced, yielding white precipitate of magnesium pyrophosphate in the reaction mixture. Judging the presence or absence of this white precipitate allows easy distinction of whether nucleic acid was amplified by the LAMP method. Since an increase in the turbidity of the reaction mixture according to the production of precipitate correlates with the amount of DNA synthesized, real-time monitoring of the LAMP reaction was achieved by real-time measurement of turbidity. Copyright 2001 Academic Press.
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              Detection of a novel human coronavirus by real-time reverse-transcription polymerase chain reaction

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                Author and article information

                Contributors
                Journal
                J Pharm Anal
                J Pharm Anal
                Journal of Pharmaceutical Analysis
                Xi'an Jiaotong University
                2095-1779
                2214-0883
                1 March 2020
                1 March 2020
                :
                Affiliations
                [1]Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, China
                Author notes
                []Corresponding author. caisheng@ 123456zju.edu.cn
                [∗∗ ]Corresponding author. zengsu@ 123456zju.edu.cn
                Article
                S2095-1779(20)30208-2
                10.1016/j.jpha.2020.02.010
                7102540
                32292623
                274964a1-0e16-42cf-a7c1-4fe9f9f45f68
                © 2020 Xi'an Jiaotong University. Production and hosting by Elsevier B.V.

                Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

                History
                : 23 February 2020
                : 27 February 2020
                : 27 February 2020
                Categories
                Article

                coronavirus,nucleic acid detection,pcr-based methods,isothermal nucleic acid amplification-based methods,microarray-based methods

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