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      Development of sandwich electrochemiluminescence immunosensor for COVID-19 diagnosis by SARS-CoV-2 Spike protein detection based on Au@BSA-Luminol nanocomposites

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          Abstract

          Coronavirus disease (COVID-19) is a new and highly contagious disease posing a threat to global public health and wreaking havoc around the world. It's caused by the Coronavirus that causes severe acute respiratory syndrome (SARS-CoV-2). In the current pandemic situation, rapid and accurate SARS-CoV-2 diagnosis on a large scale is critical for early-stage diagnosis. Early detection and monitoring of viral infections can aid in controlling and preventing infection in large groups of people. Accordingly, we developed a sensitive and high-throughput sandwich electrochemiluminescence immunosensor based on antigen detection for COVID-19 diagnosis (the spike protein of SARS-CoV-2). For the spike protein of SARS-CoV-2, the ECL biosensor had a linear range of 10 ng mL -1 to 10 µg mL -1 with a limit of detection of 1.93 ng mL -1. The sandwich ECL immunosensor could be used in early clinical diagnosis due to its excellent recovery in detecting SARS-CoV-2, rapid analysis (90 min), and ease of use.

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          Magnetic beads combined with carbon black-based screen-printed electrodes for COVID-19: A reliable and miniaturized electrochemical immunosensor for SARS-CoV-2 detection in saliva

          The diffusion of novel SARS-CoV-2 coronavirus over the world generated COVID-19 pandemic event as reported by World Health Organization on March 2020. The huge issue is the high infectivity and the absence of vaccine and customised drugs allowing for hard management of this outbreak, thus a rapid and on site analysis is a need to contain the spread of COVID-19. Herein, we developed an electrochemical immunoassay for rapid and smart detection of SARS-CoV-2 coronavirus in saliva. The electrochemical assay was conceived for Spike (S) protein or Nucleocapsid (N) protein detection using magnetic beads as support of immunological chain and secondary antibody with alkaline phosphatase as immunological label. The enzymatic by-product 1-naphtol was detected using screen-printed electrodes modified with carbon black nanomaterial. The analytical features of the electrochemical immunoassay were evaluated using the standard solution of S and N protein in buffer solution and untreated saliva with a detection limit equal to 19 ng/mL and 8 ng/mL in untreated saliva, respectively for S and N protein. Its effectiveness was assessed using cultured virus in biosafety level 3 and in saliva clinical samples comparing the data using the nasopharyngeal swab specimens tested with Real-Time PCR. The agreement of the data, the low detection limit achieved, the rapid analysis (30 min), the miniaturization, and portability of the instrument combined with the easiness to use and no-invasive sampling, confer to this analytical tool high potentiality for market entry as the first highly sensitive electrochemical immunoassay for SARS-CoV-2 detection in untreated saliva.
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            Ultrasensitive supersandwich-type electrochemical sensor for SARS-CoV-2 from the infected COVID-19 patients using a smartphone

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              Entropy-Driven Amplified Electrochemiluminescence Biosensor for RdRp Gene of SARS-CoV-2 Detection with Self-Assembled DNA Tetrahedron Scaffolds

              Dependable, specific and rapid diagnostic methods for severe acute respiratory syndrome β-coronavirus (SARS-CoV-2) detection are needed to promote public health interventions for coronavirus disease 2019 (COVID-19). Herein, we have established an entropy-driven amplified electrochemiluminescence (ECL) strategy to detect the RNA-dependent RNA polymerase (RdRp) gene of SARS-CoV-2 known as RdRp-COVID which as the target for SARS-CoV-2 plays an essential role in the diagnosis of COVID-19. For the construction of the sensors, DNA tetrahedron (DT) is modified on the surface of the electrode to furnish robust and programmable scaffolds materials, upon which target DNA-participated entropy-driven amplified reaction is efficiently conducted to link the Ru(bpy)3 2+ modified S3 to the linear ssDNA at the vertex of the tetrahedron and eventually present an “ECL on” state. The rigid tetrahedral structure of the DT probe enhances the ECL intensity and avoids the cross-reactivity between single-stranded DNA, thus increasing the sensitivity of the assays. The enzyme-free entropy-driven reaction prevents the use of expensive enzyme reagents and facilitates the realization of large-scale screening of SARS-CoV-2 patients. Our DT-based ECL sensor has demonstrated significant specificity and high sensitivity for SARS-CoV-2 with a limit of detection (LOD) down to 2.67 fM. Additionally, our operational method has achieved the detection of RdRp-COVID in human serum samples, which supplies a reliable and feasible sensing platform for the clinical bioanalysis.
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                Author and article information

                Journal
                Bioelectrochemistry
                Bioelectrochemistry
                Bioelectrochemistry (Amsterdam, Netherlands)
                Elsevier B.V.
                1567-5394
                1878-562X
                25 May 2022
                25 May 2022
                : 108161
                Affiliations
                [a ]Department of Life Science Engineering, Faculty of New Sciences & Technologies, University of Tehran, Tehran, Iran
                [b ]State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China
                [c ]Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
                [d ]Center of Excellence in Electrochemistry, School of Chemistry, University of Tehran, Tehran, Iran
                [e ]Clinical Labs, Farhikhtegan Hospital, Tehran, Iran
                Article
                S1567-5394(22)00112-8 108161
                10.1016/j.bioelechem.2022.108161
                9130314
                35659688
                e9d1fdf3-00c4-4325-a765-4bbbfda1bc85
                © 2022 Elsevier B.V. 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 February 2022
                : 11 April 2022
                : 14 May 2022
                Categories
                Article

                Biochemistry
                sars-cov-2,covid-19,electrochemiluminescence,sandwich immunosensor,au@bsa-luminol
                Biochemistry
                sars-cov-2, covid-19, electrochemiluminescence, sandwich immunosensor, au@bsa-luminol

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