Development of a Rapid Focus Reduction Neutralization Test Assay for Measuring SARS-CoV-2 Neutralizing Antibodies.


Journal

Current protocols in immunology
ISSN: 1934-368X
Titre abrégé: Curr Protoc Immunol
Pays: United States
ID NLM: 9101651

Informations de publication

Date de publication:
12 2020
Historique:
entrez: 20 11 2020
pubmed: 21 11 2020
medline: 15 12 2020
Statut: ppublish

Résumé

SARS-CoV-2 is a recently emerged human coronavirus that has escalated to a pandemic. There are currently no approved vaccines for SARS-CoV-2, which causes severe respiratory illness or death. Defining the antibody response to SARS-CoV-2 will be essential for understanding disease progression, long-term immunity, and vaccine efficacy. Here we describe two methods for evaluating the neutralization capacity of SARS-CoV-2 antibodies. The basic protocol is a focus reduction neutralization test (FRNT), which involves immunostaining infected cells with a chromogen deposit readout. The alternate protocol is a modification of the FRNT that uses an infectious clone-derived SARS-CoV-2 virus expressing a fluorescent reporter. These protocols are adapted for use in a high-throughput setting, and are compatible with large-scale vaccine studies or clinical testing. © 2020 Wiley Periodicals LLC Basic Protocol: Focus reduction neutralization test Alternate Protocol: mNeonGreen-based focus reduction neutralization test (FRNT-mNG).

Identifiants

pubmed: 33215858
doi: 10.1002/cpim.116
pmc: PMC7864545
mid: NIHMS1649342
doi:

Substances chimiques

Antibodies, Neutralizing 0

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

e116

Subventions

Organisme : NIAID NIH HHS
ID : R38 AI140299
Pays : United States
Organisme : NIAID NIH HHS
ID : U19 AI090023
Pays : United States
Organisme : NIA NIH HHS
ID : R00 AG049092
Pays : United States
Organisme : NIAID NIH HHS
ID : U19 AI057266
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI127799
Pays : United States
Organisme : NIAID NIH HHS
ID : UM1 AI148684
Pays : United States
Organisme : NIAID NIH HHS
ID : R24 AI120942
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI148378
Pays : United States
Organisme : NIH HHS
ID : P51 OD011132
Pays : United States
Organisme : NIAID NIH HHS
ID : UM1 AI148576
Pays : United States

Informations de copyright

© 2020 Wiley Periodicals LLC.

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Auteurs

Abigail Vanderheiden (A)

Department of Pediatrics, Division of Infectious Disease, Emory University School of Medicine, Atlanta, Georgia.
Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia.
Yerkes National Primate Research Center, Atlanta, Georgia.

Venkata Viswanadh Edara (VV)

Department of Pediatrics, Division of Infectious Disease, Emory University School of Medicine, Atlanta, Georgia.
Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia.
Yerkes National Primate Research Center, Atlanta, Georgia.

Katharine Floyd (K)

Department of Pediatrics, Division of Infectious Disease, Emory University School of Medicine, Atlanta, Georgia.
Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia.
Yerkes National Primate Research Center, Atlanta, Georgia.

Robert C Kauffman (RC)

Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia.
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia.

Grace Mantus (G)

Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia.
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia.

Evan Anderson (E)

Department of Pediatrics, Division of Infectious Disease, Emory University School of Medicine, Atlanta, Georgia.

Nadine Rouphael (N)

Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia.
Department of Medicine, Division of Infectious Disease, Emory University School of Medicine, Atlanta, Georgia.

Sri Edupuganti (S)

Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia.
Department of Medicine, Division of Infectious Disease, Emory University School of Medicine, Atlanta, Georgia.

Pei-Yong Shi (PY)

Department of Biochemistry and Molecular Biology, The University of Texas Medical Branch, Galveston, Texas.

Vineet D Menachery (VD)

Department of Microbiology and Immunology, Institute for Human Infection and Immunity, World Reference Center for Emerging Viruses and Arboviruses, University of Texas Medical Branch, Galveston, Texas.

Jens Wrammert (J)

Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia.
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia.

Mehul S Suthar (MS)

Department of Pediatrics, Division of Infectious Disease, Emory University School of Medicine, Atlanta, Georgia.
Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia.
Yerkes National Primate Research Center, Atlanta, Georgia.

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