SARS‐CoV‐2 Seroconversion in Humans: A Detailed Protocol for a Serological Assay, Antigen Production, and Test Setup
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Open Access
- 17 April 2020
- journal article
- research article
- Published by Wiley in Current Protocols in Microbiology
- Vol. 57 (1), e100
- https://doi.org/10.1002/cpmc.100
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‐2Keywords
Funding Information
- National Institute of Allergy and Infectious Diseases (HHSN272201400008C)
This publication has 10 references indexed in Scilit:
- Cryo-EM structure of the 2019-nCoV spike in the prefusion conformationScience, 2020
- A Novel Coronavirus from Patients with Pneumonia in China, 2019The New England Journal of Medicine, 2020
- A new coronavirus associated with human respiratory disease in ChinaNature, 2020
- Molecular Diagnosis of a Novel Coronavirus (2019-nCoV) Causing an Outbreak of PneumoniaClinical Chemistry, 2020
- Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCREurosurveillance, 2020
- Potent binding of 2019 novel coronavirus spike protein by a SARS coronavirus-specific human monoclonal antibodyEmerging Microbes & Infections, 2020
- Basic Techniques in Mammalian Cell Tissue CultureCurrent Protocols in Cell Biology, 2015
- SDS‐Polyacrylamide Gel Electrophoresis (SDS‐PAGE) of ProteinsCurrent Protocols in Microbiology, 2011
- Human Monoclonal Antibody Combination against SARS Coronavirus: Synergy and Coverage of Escape MutantsPLoS Medicine, 2006
- The time course of the immune response to experimental coronavirus infection of manEpidemiology and Infection, 1990