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      A duplex-specific nuclease based electrochemical biosensor for the assay of SARS-CoV-2 RdRp RNA

      research-article
      a , b ,
      Analytical Biochemistry
      Elsevier Inc.
      SARS-CoV-2, DSN, Electrochemistry, 2-OMe-RNA

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          Abstract

          We present a method for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection based on the dual amplification effect of duplex-specific nuclease (DSN). In this scheme, we cleverly employed a 2-OMe-RNA modified DNA to prevent hairpin nucleic acid from being digested by DSN. The target RNA and 2-OMe-RNA are released when DSN cleaves just the double-stranded RNA/hairpin nucleic acid DNA. The target RNA then forms a circular reaction when it hybridizes with another hairpin nucleic acid. Simultaneously, the released target 2-OMe-RNA turns on the hairpin DNA2 on the electrode surface, and when the DSN cleaves the DNA in the hairpin DNA2/2-OMe-RNA duplex, the 2-OMe-RNA is released and hybridized with the other hairpin DNA2. The hairpin DNA2 on the electrode surface is split off after many cycles, exposing the gold electrode surface. As a consequence, there is more K 4[Fe(CN) 6]/K 3[Fe(CN) 6] redox near to the electrode surface, and the electrochemical signal increases. As a result, the change in electrochemical signal may be used to calculate the quantity of RNA that has to be measured. The protocol has good sensitivity in the detection of SARS-CoV-2: the detection limit reached 21.69 aM. This protocol provides an effective solution for the highly sensitive screening of SARS-CoV-2.

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          CRISPR-Cas12–based detection of SARS-CoV-2

          An outbreak of betacoronavirus SARS-CoV-2 began in Wuhan, China in December 2019. COVID-19, the disease associated with infection, rapidly spread to produce a global pandemic. We report development of a rapid (<40 min), easy-to-implement and accurate CRISPR-Cas12-based lateral flow assay for detection of SARS-CoV-2 from respiratory swab RNA extracts. We validated our method using contrived reference samples and clinical samples from US patients, including 36 patients with COVID-19 infection and 42 patients with other viral respiratory infections. Our CRISPR-based DETECTR assay provides a visual and faster alternative to the US CDC SARS-CoV-2 real-time RT-PCR assay, with 95% positive predictive agreement and 100% negative predictive agreement.. SARS-CoV-2 in patient samples is detected in under an hour using a CRISPR-based lateral flow assay.
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            Dual-Functional Plasmonic Photothermal Biosensors for Highly Accurate Severe Acute Respiratory Syndrome Coronavirus 2 Detection

            The ongoing outbreak of the novel coronavirus disease (COVID-19) has spread globally and poses a threat to public health in more than 200 countries. Reliable laboratory diagnosis of the disease has been one of the foremost priorities for promoting public health interventions. The routinely used reverse transcription polymerase chain reaction (RT-PCR) is currently the reference method for COVID-19 diagnosis. However, it also reported a number of false-positive or -negative cases, especially in the early stages of the novel virus outbreak. In this work, a dual-functional plasmonic biosensor combining the plasmonic photothermal (PPT) effect and localized surface plasmon resonance (LSPR) sensing transduction provides an alternative and promising solution for the clinical COVID-19 diagnosis. The two-dimensional gold nanoislands (AuNIs) functionalized with complementary DNA receptors can perform a sensitive detection of the selected sequences from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) through nucleic acid hybridization. For better sensing performance, the thermoplasmonic heat is generated on the same AuNIs chip when illuminated at their plasmonic resonance frequency. The localized PPT heat is capable to elevate the in situ hybridization temperature and facilitate the accurate discrimination of two similar gene sequences. Our dual-functional LSPR biosensor exhibits a high sensitivity toward the selected SARS-CoV-2 sequences with a lower detection limit down to the concentration of 0.22 pM and allows precise detection of the specific target in a multigene mixture. This study gains insight into the thermoplasmonic enhancement and its applicability in the nucleic acid tests and viral disease diagnosis.
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              Homogeneous circle-to-circle amplification for real-time optomagnetic detection of SARS-CoV-2 RdRp coding sequence

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                Author and article information

                Journal
                Anal Biochem
                Anal Biochem
                Analytical Biochemistry
                Elsevier Inc.
                0003-2697
                1096-0309
                21 November 2022
                15 January 2023
                21 November 2022
                : 661
                : 114983
                Affiliations
                [a ]Department of Laboratory Medicine, Jiangyin Hospital of Traditional Chinese Medicine (Jiangyin Hospital Affiliated to Nanjing University of Chinese Medicine), Jiangyin, Jiangsu, China
                [b ]Department of Laboratory Medicine, Jiangyin Fifth People's Hospital, Jiangyin, Jiangsu, China
                Author notes
                []Corresponding author.
                Article
                S0003-2697(22)00443-2 114983
                10.1016/j.ab.2022.114983
                9676167
                74259d96-a569-4a0e-a203-f0d3fd300a2a
                © 2022 Elsevier Inc. All rights reserved.

                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
                : 12 September 2022
                : 26 October 2022
                : 8 November 2022
                Categories
                Article

                Biochemistry
                sars-cov-2,dsn,electrochemistry,2-ome-rna
                Biochemistry
                sars-cov-2, dsn, electrochemistry, 2-ome-rna

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