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An enzyme-based immunodetection assay to quantify SARS-CoV-2 infection.
Antiviral Res. 2020 09; 181:104882.AR

Abstract

SARS-CoV-2 is a novel pandemic coronavirus that caused a global health and economic crisis. The development of efficient drugs and vaccines against COVID-19 requires detailed knowledge about SARS-CoV-2 biology. Several techniques to detect SARS-CoV-2 infection have been established, mainly based on counting infected cells by staining plaques or foci, or by quantifying the viral genome by PCR. These methods are laborious, time-consuming and expensive and therefore not suitable for a high sample throughput or rapid diagnostics. We here report a novel enzyme-based immunodetection assay that directly quantifies the amount of de novo synthesized viral spike protein within fixed and permeabilized cells. This in-cell ELISA enables a rapid and quantitative detection of SARS-CoV-2 infection in microtiter format, regardless of the virus isolate or target cell culture. It follows the established method of performing ELISA assays and does not require expensive instrumentation. Utilization of the in-cell ELISA allows to e.g. determine TCID50 of virus stocks, antiviral efficiencies (IC50 values) of drugs or neutralizing activity of sera. Thus, the in-cell spike ELISA represents a promising alternative to study SARS-CoV-2 infection and inhibition and may facilitate future research.

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Authors+Show Affiliations

Conzelmann C
Institute of Molecular Virology, Ulm University Medical Center, 89081, Ulm, Germany.
Gilg A
Institute of Molecular Virology, Ulm University Medical Center, 89081, Ulm, Germany.
Groβ R
Institute of Molecular Virology, Ulm University Medical Center, 89081, Ulm, Germany.
Schütz D
Institute of Molecular Virology, Ulm University Medical Center, 89081, Ulm, Germany.
Preising N
Core Facility Functional Peptidomics, Ulm University Medical Center, 89081, Ulm, Germany.
Ständker L
Core Facility Functional Peptidomics, Ulm University Medical Center, 89081, Ulm, Germany.
Jahrsdörfer B
Institute for Transfusion Medicine, Ulm University, 89081, Ulm, Germany; Institute for Clinical Transfusion Medicine and Immunogenetics Ulm, German Red Cross Blood Services Baden-Württemberg-Hessen and University Hospital Ulm, 89081, Ulm, Germany.
Schrezenmeier H
Institute for Transfusion Medicine, Ulm University, 89081, Ulm, Germany; Institute for Clinical Transfusion Medicine and Immunogenetics Ulm, German Red Cross Blood Services Baden-Württemberg-Hessen and University Hospital Ulm, 89081, Ulm, Germany.
Sparrer KMJ
Institute of Molecular Virology, Ulm University Medical Center, 89081, Ulm, Germany.
Stamminger T
Institute of Virology, Ulm University Medical Center, 89081, Ulm, Germany.
Stenger S
Institute of Medical Microbiology and Hygiene, Ulm University Medical Center, 89081, Ulm, Germany.
Münch J
Institute of Molecular Virology, Ulm University Medical Center, 89081, Ulm, Germany; Core Facility Functional Peptidomics, Ulm University Medical Center, 89081, Ulm, Germany.
Müller JA
Institute of Molecular Virology, Ulm University Medical Center, 89081, Ulm, Germany. Electronic address: Janis.mueller@uni-ulm.de.

MeSH

AnimalsAntibodies, ViralBetacoronavirusCOVID-19Chlorocebus aethiopsCoronavirus InfectionsEnzyme-Linked Immunosorbent AssayHumansPandemicsPneumonia, ViralSARS-CoV-2Severe Acute Respiratory SyndromeSpike Glycoprotein, CoronavirusVero CellsViral Plaque Assay

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

32738255

Clinical Trial Links

Clinical trial which this article describes

Citation

Conzelmann, Carina, et al. "An Enzyme-based Immunodetection Assay to Quantify SARS-CoV-2 Infection." Antiviral Research, vol. 181, 2020, p. 104882.
Conzelmann C, Gilg A, Groβ R, et al. An enzyme-based immunodetection assay to quantify SARS-CoV-2 infection. Antiviral Res. 2020;181:104882.
Conzelmann, C., Gilg, A., Groβ, R., Schütz, D., Preising, N., Ständker, L., Jahrsdörfer, B., Schrezenmeier, H., Sparrer, K. M. J., Stamminger, T., Stenger, S., Münch, J., & Müller, J. A. (2020). An enzyme-based immunodetection assay to quantify SARS-CoV-2 infection. Antiviral Research, 181, 104882. https://doi.org/10.1016/j.antiviral.2020.104882
Conzelmann C, et al. An Enzyme-based Immunodetection Assay to Quantify SARS-CoV-2 Infection. Antiviral Res. 2020;181:104882. PubMed PMID: 32738255.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - An enzyme-based immunodetection assay to quantify SARS-CoV-2 infection. AU - Conzelmann,Carina, AU - Gilg,Andrea, AU - Groβ,Rüdiger, AU - Schütz,Desiree, AU - Preising,Nico, AU - Ständker,Ludger, AU - Jahrsdörfer,Bernd, AU - Schrezenmeier,Hubert, AU - Sparrer,Konstantin M J, AU - Stamminger,Thomas, AU - Stenger,Steffen, AU - Münch,Jan, AU - Müller,Janis A, Y1 - 2020/07/29/ PY - 2020/06/14/received PY - 2020/07/12/revised PY - 2020/07/14/accepted PY - 2020/8/2/pubmed PY - 2020/9/8/medline PY - 2020/8/2/entrez KW - Antiviral testing KW - Drug screening KW - In-cell ELISA KW - Neutralization KW - SARS-CoV-2 SP - 104882 EP - 104882 JF - Antiviral research JO - Antiviral Res VL - 181 N2 - SARS-CoV-2 is a novel pandemic coronavirus that caused a global health and economic crisis. The development of efficient drugs and vaccines against COVID-19 requires detailed knowledge about SARS-CoV-2 biology. Several techniques to detect SARS-CoV-2 infection have been established, mainly based on counting infected cells by staining plaques or foci, or by quantifying the viral genome by PCR. These methods are laborious, time-consuming and expensive and therefore not suitable for a high sample throughput or rapid diagnostics. We here report a novel enzyme-based immunodetection assay that directly quantifies the amount of de novo synthesized viral spike protein within fixed and permeabilized cells. This in-cell ELISA enables a rapid and quantitative detection of SARS-CoV-2 infection in microtiter format, regardless of the virus isolate or target cell culture. It follows the established method of performing ELISA assays and does not require expensive instrumentation. Utilization of the in-cell ELISA allows to e.g. determine TCID50 of virus stocks, antiviral efficiencies (IC50 values) of drugs or neutralizing activity of sera. Thus, the in-cell spike ELISA represents a promising alternative to study SARS-CoV-2 infection and inhibition and may facilitate future research. SN - 1872-9096 UR - https://www.unboundmedicine.com/medline/citation/32738255/An_enzyme_based_immunodetection_assay_to_quantify_SARS_CoV_2_infection_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0166-3542(20)30296-5 DB - PRIME DP - Unbound Medicine ER -