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SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup.
Curr Protoc Microbiol. 2020 06; 57(1):e100.CP

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.

Authors+Show Affiliations

Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York. Graduate School of Biomedical Sciences, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York. Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York. Graduate School of Biomedical Sciences, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York. Graduate School of Biomedical Sciences, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York. Graduate School of Biomedical Sciences, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York. Graduate School of Biomedical Sciences, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York. Global Health Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York. Division of Infectious Diseases, Department of Medicine, Icahn School of Medicine at Mount Sinai, New York, New York.Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

32302069

Citation

Stadlbauer, Daniel, et al. "SARS-CoV-2 Seroconversion in Humans: a Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup." Current Protocols in Microbiology, vol. 57, no. 1, 2020, pp. e100.
Stadlbauer D, Amanat F, Chromikova V, et al. SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup. Curr Protoc Microbiol. 2020;57(1):e100.
Stadlbauer, D., Amanat, F., Chromikova, V., Jiang, K., Strohmeier, S., Arunkumar, G. A., Tan, J., Bhavsar, D., Capuano, C., Kirkpatrick, E., Meade, P., Brito, R. N., Teo, C., McMahon, M., Simon, V., & Krammer, F. (2020). SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup. Current Protocols in Microbiology, 57(1), e100. https://doi.org/10.1002/cpmc.100
Stadlbauer D, et al. SARS-CoV-2 Seroconversion in Humans: a Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup. Curr Protoc Microbiol. 2020;57(1):e100. PubMed PMID: 32302069.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - SARS-CoV-2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup. AU - Stadlbauer,Daniel, AU - Amanat,Fatima, AU - Chromikova,Veronika, AU - Jiang,Kaijun, AU - Strohmeier,Shirin, AU - Arunkumar,Guha Asthagiri, AU - Tan,Jessica, AU - Bhavsar,Disha, AU - Capuano,Christina, AU - Kirkpatrick,Ericka, AU - Meade,Philip, AU - Brito,Ruhi Nichalle, AU - Teo,Catherine, AU - McMahon,Meagan, AU - Simon,Viviana, AU - Krammer,Florian, PY - 2020/4/18/entrez PY - 2020/4/18/pubmed PY - 2020/4/23/medline KW - COVID19 KW - ELISA KW - SARS-CoV-2 KW - protein expression KW - serological assay SP - e100 EP - e100 JF - Current protocols in microbiology JO - Curr Protoc Microbiol VL - 57 IS - 1 N2 - 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. SN - 1934-8533 UR - https://www.unboundmedicine.com/medline/citation/32302069/SARS_CoV_2_Seroconversion_in_Humans:_A_Detailed_Protocol_for_a_Serological_Assay_Antigen_Production_and_Test_Setup_ L2 - https://doi.org/10.1002/cpmc.100 DB - PRIME DP - Unbound Medicine ER -