SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup.
Identifieur interne : 000278 ( 2020/Analysis ); précédent : 000277; suivant : 000279SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup.
Auteurs : Daniel Stadlbauer [États-Unis] ; Fatima Amanat [États-Unis] ; Veronika Chromikova [États-Unis] ; Kaijun Jiang [États-Unis] ; Shirin Strohmeier [États-Unis] ; Guha Asthagiri Arunkumar [États-Unis] ; Jessica Tan [États-Unis] ; Disha Bhavsar [États-Unis] ; Christina Capuano [États-Unis] ; Ericka Kirkpatrick [États-Unis] ; Philip Meade [États-Unis] ; Ruhi Nichalle Brito [États-Unis] ; Catherine Teo [États-Unis] ; Meagan Mcmahon [États-Unis] ; Viviana Simon [États-Unis] ; Florian Krammer [États-Unis]Source :
- Current protocols in microbiology [ 1934-8533 ] ; 2020.
Abstract
In late 2019, cases of atypical pneumonia were detected in China. The etiological agent was quickly identified as a betacoronavirus (named SARS-CoV-2), which has since caused a pandemic. Several methods allowing for the specific detection of viral nucleic acids have been established, but these only allow detection of the virus during a short period of time, generally during acute infection. Serological assays are urgently needed to conduct serosurveys, to understand the antibody responses mounted in response to the virus, and to identify individuals who are potentially immune to re-infection. Here we describe a detailed protocol for expression of antigens derived from the spike protein of SARS-CoV-2 that can serve as a substrate for immunological assays, as well as a two-stage serological enzyme-linked immunosorbent assay (ELISA). These assays can be used for research studies and for testing in clinical laboratories. © 2020 The Authors. Basic Protocol 1: Mammalian cell transfection and protein purification Basic Protocol 2: A two-stage ELISA for high-throughput screening of human serum samples for antibodies binding to the spike protein of SARS-CoV-2.
DOI: 10.1002/cpmc.100
PubMed: 32302069
Affiliations:
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pubmed:32302069Le document en format XML
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xml:lang="en">SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup.</title><author><name sortKey="Stadlbauer, Daniel" sort="Stadlbauer, Daniel" uniqKey="Stadlbauer D" first="Daniel" last="Stadlbauer">Daniel Stadlbauer</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author><author><name sortKey="Amanat, Fatima" sort="Amanat, Fatima" uniqKey="Amanat F" first="Fatima" last="Amanat">Fatima Amanat</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author><author><name sortKey="Chromikova, Veronika" sort="Chromikova, Veronika" uniqKey="Chromikova V" first="Veronika" last="Chromikova">Veronika Chromikova</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author><author><name sortKey="Jiang, Kaijun" sort="Jiang, Kaijun" uniqKey="Jiang K" first="Kaijun" last="Jiang">Kaijun Jiang</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author><author><name sortKey="Strohmeier, Shirin" sort="Strohmeier, Shirin" uniqKey="Strohmeier S" first="Shirin" last="Strohmeier">Shirin Strohmeier</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author><author><name sortKey="Arunkumar, Guha Asthagiri" sort="Arunkumar, Guha Asthagiri" uniqKey="Arunkumar G" first="Guha Asthagiri" last="Arunkumar">Guha Asthagiri Arunkumar</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author><author><name sortKey="Tan, Jessica" sort="Tan, Jessica" uniqKey="Tan J" first="Jessica" last="Tan">Jessica Tan</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author><author><name sortKey="Bhavsar, Disha" sort="Bhavsar, Disha" uniqKey="Bhavsar D" first="Disha" last="Bhavsar">Disha Bhavsar</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author><author><name sortKey="Capuano, Christina" sort="Capuano, Christina" uniqKey="Capuano C" first="Christina" last="Capuano">Christina Capuano</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author><author><name sortKey="Kirkpatrick, Ericka" sort="Kirkpatrick, Ericka" uniqKey="Kirkpatrick E" first="Ericka" last="Kirkpatrick">Ericka Kirkpatrick</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author><author><name sortKey="Meade, Philip" sort="Meade, Philip" uniqKey="Meade P" first="Philip" last="Meade">Philip Meade</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author><author><name sortKey="Brito, Ruhi Nichalle" sort="Brito, Ruhi Nichalle" uniqKey="Brito R" first="Ruhi Nichalle" last="Brito">Ruhi Nichalle Brito</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author><author><name sortKey="Teo, Catherine" sort="Teo, Catherine" uniqKey="Teo C" first="Catherine" last="Teo">Catherine Teo</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country 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Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author><author><name sortKey="Krammer, Florian" sort="Krammer, Florian" uniqKey="Krammer F" first="Florian" last="Krammer">Florian Krammer</name><affiliation wicri:level="2"><nlm:affiliation>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York</wicri:cityArea></affiliation></author></analytic><series><title level="j">Current protocols in microbiology</title><idno type="eISSN">1934-8533</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">In late 2019, cases of atypical pneumonia were detected in China. The etiological agent was quickly identified as a betacoronavirus (named SARS-CoV-2), which has since caused a pandemic. Several methods allowing for the specific detection of viral nucleic acids have been established, but these only allow detection of the virus during a short period of time, generally during acute infection. Serological assays are urgently needed to conduct serosurveys, to understand the antibody responses mounted in response to the virus, and to identify individuals who are potentially immune to re-infection. Here we describe a detailed protocol for expression of antigens derived from the spike protein of SARS-CoV-2 that can serve as a substrate for immunological assays, as well as a two-stage serological enzyme-linked immunosorbent assay (ELISA). These assays can be used for research studies and for testing in clinical laboratories. © 2020 The Authors. Basic Protocol 1: Mammalian cell transfection and protein purification Basic Protocol 2: A two-stage ELISA for high-throughput screening of human serum samples for antibodies binding to the spike protein of SARS-CoV-2.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>État de New York</li></region></list><tree><country name="États-Unis"><region name="État de New York"><name sortKey="Stadlbauer, Daniel" sort="Stadlbauer, Daniel" uniqKey="Stadlbauer D" first="Daniel" last="Stadlbauer">Daniel Stadlbauer</name></region><name sortKey="Amanat, Fatima" sort="Amanat, Fatima" uniqKey="Amanat F" first="Fatima" last="Amanat">Fatima Amanat</name><name sortKey="Arunkumar, Guha Asthagiri" sort="Arunkumar, Guha Asthagiri" uniqKey="Arunkumar G" first="Guha Asthagiri" last="Arunkumar">Guha Asthagiri Arunkumar</name><name sortKey="Bhavsar, Disha" sort="Bhavsar, Disha" uniqKey="Bhavsar D" first="Disha" last="Bhavsar">Disha Bhavsar</name><name sortKey="Brito, Ruhi Nichalle" sort="Brito, Ruhi Nichalle" uniqKey="Brito R" first="Ruhi Nichalle" last="Brito">Ruhi Nichalle Brito</name><name sortKey="Capuano, Christina" sort="Capuano, Christina" uniqKey="Capuano C" first="Christina" last="Capuano">Christina Capuano</name><name sortKey="Chromikova, Veronika" sort="Chromikova, Veronika" uniqKey="Chromikova V" first="Veronika" last="Chromikova">Veronika Chromikova</name><name sortKey="Jiang, Kaijun" sort="Jiang, Kaijun" uniqKey="Jiang K" first="Kaijun" last="Jiang">Kaijun Jiang</name><name sortKey="Kirkpatrick, Ericka" sort="Kirkpatrick, Ericka" uniqKey="Kirkpatrick E" first="Ericka" last="Kirkpatrick">Ericka Kirkpatrick</name><name sortKey="Krammer, Florian" sort="Krammer, Florian" uniqKey="Krammer F" first="Florian" last="Krammer">Florian Krammer</name><name sortKey="Mcmahon, Meagan" sort="Mcmahon, Meagan" uniqKey="Mcmahon M" first="Meagan" last="Mcmahon">Meagan Mcmahon</name><name sortKey="Meade, Philip" sort="Meade, Philip" uniqKey="Meade P" first="Philip" last="Meade">Philip Meade</name><name sortKey="Simon, Viviana" sort="Simon, Viviana" uniqKey="Simon V" first="Viviana" last="Simon">Viviana Simon</name><name sortKey="Strohmeier, Shirin" sort="Strohmeier, Shirin" uniqKey="Strohmeier S" first="Shirin" last="Strohmeier">Shirin Strohmeier</name><name sortKey="Tan, Jessica" sort="Tan, Jessica" uniqKey="Tan J" first="Jessica" last="Tan">Jessica Tan</name><name sortKey="Teo, Catherine" sort="Teo, Catherine" uniqKey="Teo C" first="Catherine" last="Teo">Catherine Teo</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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