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      Evaluation of a multiplexed coronavirus antigen array for detection of SARS-CoV-2 specific IgG in COVID-19 convalescent plasma

      research-article
      a , b , a , b , c , c , b , d , b , d , e , e , a , b , *
      Journal of Immunological Methods
      Published by Elsevier B.V.
      COVID-19, SARS-CoV-2, Antigen array, Spike antigen, SARS-CoV-2 IgG, COVID-19, Coronavirus Disease 2019, SARS-CoV-2, Severe Acute Respiratory Syndrome Coronavirus-2, CoV, Coronavirus, S, Spike protein, RBD, Receptor Binding Domain, ELISA, Enzyme Linked Immunosorbent Assay, FDA, Food and Drug Administration, EUA, Emergency Use Authorization, CCP, COVID-19 Convalescent Plasma, RPP, Respiratory Pathogen Panel, EDI, Epitope Diagnostic Inc, HRP, Horseradish peroxidase, TMB, Tetramethyl Benzidine, FRNT, Focus Reduction Neutralizing Antibody Test, OD, Optical Density

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          Abstract

          Mitigation of the COVID-19 pandemic requires an understanding of the antibody response to SARS-CoV-2. However, throughout the development of SARS-CoV-2 IgG antibody assays during the past year, cross-reactivity to other coronaviruses remained a question. To address these issues, we evaluated IgG in COVID-19 convalescent plasma samples for reactivity against three SARS-CoV-2 antigens including full-length spike, receptor binding domain, and the proximal extracellular fusion domain, and spike antigens from other coronaviruses (SARS-CoV, MERS-CoV, hCoV-HKU1, hCoV-OC43, hCoV-229E and hCoV-NL63) using the VaxArray Coronavirus SeroAssay which is a multiplexed antigen assay developed by InDevR Inc. These results were compared to two commercial SARS-CoV-2 IgG ELISAs targeting either the SARS-CoV-2 nucleocapsid or spike antigens and a live virus focus reduction neutralizing antibody test (FRNT). The VaxArray platform showed high specificity for detection of SARS-CoV-2 IgG, evident from lack of reactivity to SARS-CoV-2 antigens despite significant reactivity to endemic coronavirus antigens in pre-pandemic samples. SARS-CoV-2 IgG positive samples reacted weakly to SARS-CoV spike but not to MERS-CoV. While the VaxArray platform had overall comparable results to the spike and nucleocapsid IgG ELISAs, results were more similar to the spike antigen ELISA and the platform displayed a higher sensitivity and specificity than both ELISAs. Samples with FRNT titers below 1/23 reported negative on VaxArray, while positive samples on VaxArray had significantly higher neutralizing antibody titers. These results suggest that the VaxArray Coronavirus SeroAssay performs with high sensitivity and specificity for the detection of SARS-CoV-2 IgG, and positive results on the platform indicate SARS-CoV-2 neutralizing activity.

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          Emerging coronaviruses: Genome structure, replication, and pathogenesis

          Abstract The recent emergence of a novel coronavirus (2019‐nCoV), which is causing an outbreak of unusual viral pneumonia in patients in Wuhan, a central city in China, is another warning of the risk of CoVs posed to public health. In this minireview, we provide a brief introduction of the general features of CoVs and describe diseases caused by different CoVs in humans and animals. This review will help understand the biology and potential risk of CoVs that exist in richness in wildlife such as bats.
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            Treatment of 5 Critically Ill Patients With COVID-19 With Convalescent Plasma

            Could administration of convalescent plasma transfusion be beneficial in the treatment of critically ill patients with coronavirus disease 2019 (COVID-19)? In this uncontrolled case series of 5 critically ill patients with COVID-19 and acute respiratory distress syndrome (ARDS), administration of convalescent plasma containing neutralizing antibody was followed by an improvement in clinical status. These preliminary findings raise the possibility that convalescent plasma transfusion may be helpful in the treatment of critically ill patients with COVID-19 and ARDS, but this approach requires evaluation in randomized clinical trials. Coronavirus disease 2019 (COVID-19) is a pandemic with no specific therapeutic agents and substantial mortality. It is critical to find new treatments. To determine whether convalescent plasma transfusion may be beneficial in the treatment of critically ill patients with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Case series of 5 critically ill patients with laboratory-confirmed COVID-19 and acute respiratory distress syndrome (ARDS) who met the following criteria: severe pneumonia with rapid progression and continuously high viral load despite antiviral treatment; P ao 2 /F io 2 <300; and mechanical ventilation. All 5 were treated with convalescent plasma transfusion. The study was conducted at the infectious disease department, Shenzhen Third People's Hospital in Shenzhen, China, from January 20, 2020, to March 25, 2020; final date of follow-up was March 25, 2020. Clinical outcomes were compared before and after convalescent plasma transfusion. Patients received transfusion with convalescent plasma with a SARS-CoV-2–specific antibody (IgG) binding titer greater than 1:1000 (end point dilution titer, by enzyme-linked immunosorbent assay [ELISA]) and a neutralization titer greater than 40 (end point dilution titer) that had been obtained from 5 patients who recovered from COVID-19. Convalescent plasma was administered between 10 and 22 days after admission. Changes of body temperature, Sequential Organ Failure Assessment (SOFA) score (range 0-24, with higher scores indicating more severe illness), P ao 2 /F io 2 , viral load, serum antibody titer, routine blood biochemical index, ARDS, and ventilatory and extracorporeal membrane oxygenation (ECMO) supports before and after convalescent plasma transfusion. All 5 patients (age range, 36-65 years; 2 women) were receiving mechanical ventilation at the time of treatment and all had received antiviral agents and methylprednisolone. Following plasma transfusion, body temperature normalized within 3 days in 4 of 5 patients, the SOFA score decreased, and P ao 2 /F io 2 increased within 12 days (range, 172-276 before and 284-366 after). Viral loads also decreased and became negative within 12 days after the transfusion, and SARS-CoV-2–specific ELISA and neutralizing antibody titers increased following the transfusion (range, 40-60 before and 80-320 on day 7). ARDS resolved in 4 patients at 12 days after transfusion, and 3 patients were weaned from mechanical ventilation within 2 weeks of treatment. Of the 5 patients, 3 have been discharged from the hospital (length of stay: 53, 51, and 55 days), and 2 are in stable condition at 37 days after transfusion. In this preliminary uncontrolled case series of 5 critically ill patients with COVID-19 and ARDS, administration of convalescent plasma containing neutralizing antibody was followed by improvement in their clinical status. The limited sample size and study design preclude a definitive statement about the potential effectiveness of this treatment, and these observations require evaluation in clinical trials. This case series describes clinical outcomes in 5 Chinese patients with laboratory-confirmed COVID-19, ARDS, and high viral loads despite antiviral treatment who were given human plasma with SARS-CoV-2 antibodies obtained from previously infected and recovered patients.
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              Severe Acute Respiratory Syndrome Coronavirus 2 from Patient with Coronavirus Disease, United States

              The etiologic agent of an outbreak of pneumonia in Wuhan, China, was identified as severe acute respiratory syndrome coronavirus 2 in January 2020. A patient in the United States was given a diagnosis of infection with this virus by the state of Washington and the US Centers for Disease Control and Prevention on January 20, 2020. We isolated virus from nasopharyngeal and oropharyngeal specimens from this patient and characterized the viral sequence, replication properties, and cell culture tropism. We found that the virus replicates to high titer in Vero-CCL81 cells and Vero E6 cells in the absence of trypsin. We also deposited the virus into 2 virus repositories, making it broadly available to the public health and research communities. We hope that open access to this reagent will expedite development of medical countermeasures.
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                Author and article information

                Journal
                J Immunol Methods
                J Immunol Methods
                Journal of Immunological Methods
                Published by Elsevier B.V.
                0022-1759
                1872-7905
                22 July 2021
                22 July 2021
                Affiliations
                [a ]Department of Pediatrics, University of Colorado School of Medicine, USA
                [b ]Children's Hospital, 13123 East 16 th Avenue, Aurora, CO 80045, USA
                [c ]Department of Immunology and Microbiology, University of Colorado School of Medicine, USA
                [d ]Department of Pathology and Laboratory Medicine, University of Colorado School of Medicine and Children's Hospital, CO, USA
                [e ]InDevR Inc., 2100 Central Ave., Suite 106, Boulder, CO 80301, USA
                Author notes
                [* ]Corresponding author at: Pediatrics, Section of Allergy and Immunology, University of Colorado School of Medicine, Translational and Diagnostic Immunology Laboratory, Children's Hospital, 13123 East 16th Avenue, B518, Aurora, CO 80045, USA.
                Article
                S0022-1759(21)00149-6 113104
                10.1016/j.jim.2021.113104
                8297968
                34303688
                59ed4a54-c843-43c7-9fa3-85df914749e2
                © 2021 Published 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.

                Categories
                Article

                Immunology
                covid-19,sars-cov-2,antigen array,spike antigen,sars-cov-2 igg,covid-19, coronavirus disease 2019,sars-cov-2, severe acute respiratory syndrome coronavirus-2,cov, coronavirus,s, spike protein,rbd, receptor binding domain,elisa, enzyme linked immunosorbent assay,fda, food and drug administration,eua, emergency use authorization,ccp, covid-19 convalescent plasma,rpp, respiratory pathogen panel,edi, epitope diagnostic inc,hrp, horseradish peroxidase,tmb, tetramethyl benzidine,frnt, focus reduction neutralizing antibody test,od, optical density

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