Neutralization assay with SARS-CoV-1 and SARS-CoV-2 spike pseudotyped murine leukemia virions
Abstract Background Virus neutralization by antibodies is an important prognostic factor in many viral diseases. To easily and rapidly measure titers of neutralizing antibodies in serum or plasma, we developed pseudovirion particles composed of the spike glycoprotein of SARS-CoV-2 incorporated onto...
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doaj-23bf90048a4649b098183e1247d1a8c62021-01-10T12:38:22ZengBMCVirology Journal1743-422X2021-01-011811610.1186/s12985-020-01472-1Neutralization assay with SARS-CoV-1 and SARS-CoV-2 spike pseudotyped murine leukemia virionsYue Zheng0Erin T. Larragoite1Elizabeth S. C. P. Williams2Juan Lama3Isabel Cisneros4Julio C. Delgado5Patricia Slev6Jenna Rychert7Emily A. Innis8Mayte Coiras9Matthew T. Rondina10Adam M. Spivak11Vicente Planelles12Department of Pathology, University of Utah School of MedicineDepartment of Pathology, University of Utah School of MedicineDepartment of Pathology, University of Utah School of MedicineRetroVirox, Inc.RetroVirox, Inc.Department of Pathology, University of Utah School of MedicineDepartment of Pathology, University of Utah School of MedicineAssociated Regional and University Pathologists (ARUP) LaboratoriesDepartment of Pathology, University of Utah School of MedicineAIDS Immunopathology Unit, National Center of Microbiology (CNM), Instituto de Salud Carlos IIIDepartment of Human Genetics, University of Utah School of MedicineDepartment of Medicine, University of Utah School of MedicineDepartment of Pathology, University of Utah School of MedicineAbstract Background Virus neutralization by antibodies is an important prognostic factor in many viral diseases. To easily and rapidly measure titers of neutralizing antibodies in serum or plasma, we developed pseudovirion particles composed of the spike glycoprotein of SARS-CoV-2 incorporated onto murine leukemia virus capsids and a modified minimal murine leukemia virus genome encoding firefly luciferase. This assay design is intended for use in laboratories with biocontainment level 2 and therefore circumvents the need for the biocontainment level 3 that would be required for replication-competent SARS-CoV-2 virus. To validate the pseudovirion assay, we set up comparisons with other available antibody tests including those from Abbott, Euroimmun and Siemens, using archived, known samples. Results 11 out of 12 SARS-CoV-2-infected patient serum samples showed neutralizing activity against SARS-CoV-2-spike pseudotyped MLV viruses, with neutralizing titers-50 (NT50) that ranged from 1:25 to 1:1,417. Five historical samples from patients hospitalized for severe influenza infection in 2016 tested negative in the neutralization assay (NT50 < 25). Three serum samples with high neutralizing activity against SARS-CoV-2/MLV pseudoviruses showed no detectable neutralizing activity (NT50 < 25) against SARS-CoV-1/MLV pseudovirions. We also compared the semiquantitative Siemens SARS-CoV-2 IgG test, which measures binding of IgG to recombinantly expressed receptor binding domain of SARS-CoV-2 spike glycoprotein with the neutralization titers obtained in the pseudovirion assay and the results show high concordance between the two tests (R2 = 0.9344). Conclusions SARS-CoV-2 spike/MLV pseudovirions provide a practical means of assessing neutralizing activity of antibodies in serum or plasma from infected patients under laboratory conditions consistent with biocontainment level 2. This assay offers promise also in evaluating immunogenicity of spike glycoprotein-based candidate vaccines in the near future.https://doi.org/10.1186/s12985-020-01472-1COVID-19CoronavirusSARSSARS-CoV-2Neutralization assayPseudotyped virus |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yue Zheng Erin T. Larragoite Elizabeth S. C. P. Williams Juan Lama Isabel Cisneros Julio C. Delgado Patricia Slev Jenna Rychert Emily A. Innis Mayte Coiras Matthew T. Rondina Adam M. Spivak Vicente Planelles |
spellingShingle |
Yue Zheng Erin T. Larragoite Elizabeth S. C. P. Williams Juan Lama Isabel Cisneros Julio C. Delgado Patricia Slev Jenna Rychert Emily A. Innis Mayte Coiras Matthew T. Rondina Adam M. Spivak Vicente Planelles Neutralization assay with SARS-CoV-1 and SARS-CoV-2 spike pseudotyped murine leukemia virions Virology Journal COVID-19 Coronavirus SARS SARS-CoV-2 Neutralization assay Pseudotyped virus |
author_facet |
Yue Zheng Erin T. Larragoite Elizabeth S. C. P. Williams Juan Lama Isabel Cisneros Julio C. Delgado Patricia Slev Jenna Rychert Emily A. Innis Mayte Coiras Matthew T. Rondina Adam M. Spivak Vicente Planelles |
author_sort |
Yue Zheng |
title |
Neutralization assay with SARS-CoV-1 and SARS-CoV-2 spike pseudotyped murine leukemia virions |
title_short |
Neutralization assay with SARS-CoV-1 and SARS-CoV-2 spike pseudotyped murine leukemia virions |
title_full |
Neutralization assay with SARS-CoV-1 and SARS-CoV-2 spike pseudotyped murine leukemia virions |
title_fullStr |
Neutralization assay with SARS-CoV-1 and SARS-CoV-2 spike pseudotyped murine leukemia virions |
title_full_unstemmed |
Neutralization assay with SARS-CoV-1 and SARS-CoV-2 spike pseudotyped murine leukemia virions |
title_sort |
neutralization assay with sars-cov-1 and sars-cov-2 spike pseudotyped murine leukemia virions |
publisher |
BMC |
series |
Virology Journal |
issn |
1743-422X |
publishDate |
2021-01-01 |
description |
Abstract Background Virus neutralization by antibodies is an important prognostic factor in many viral diseases. To easily and rapidly measure titers of neutralizing antibodies in serum or plasma, we developed pseudovirion particles composed of the spike glycoprotein of SARS-CoV-2 incorporated onto murine leukemia virus capsids and a modified minimal murine leukemia virus genome encoding firefly luciferase. This assay design is intended for use in laboratories with biocontainment level 2 and therefore circumvents the need for the biocontainment level 3 that would be required for replication-competent SARS-CoV-2 virus. To validate the pseudovirion assay, we set up comparisons with other available antibody tests including those from Abbott, Euroimmun and Siemens, using archived, known samples. Results 11 out of 12 SARS-CoV-2-infected patient serum samples showed neutralizing activity against SARS-CoV-2-spike pseudotyped MLV viruses, with neutralizing titers-50 (NT50) that ranged from 1:25 to 1:1,417. Five historical samples from patients hospitalized for severe influenza infection in 2016 tested negative in the neutralization assay (NT50 < 25). Three serum samples with high neutralizing activity against SARS-CoV-2/MLV pseudoviruses showed no detectable neutralizing activity (NT50 < 25) against SARS-CoV-1/MLV pseudovirions. We also compared the semiquantitative Siemens SARS-CoV-2 IgG test, which measures binding of IgG to recombinantly expressed receptor binding domain of SARS-CoV-2 spike glycoprotein with the neutralization titers obtained in the pseudovirion assay and the results show high concordance between the two tests (R2 = 0.9344). Conclusions SARS-CoV-2 spike/MLV pseudovirions provide a practical means of assessing neutralizing activity of antibodies in serum or plasma from infected patients under laboratory conditions consistent with biocontainment level 2. This assay offers promise also in evaluating immunogenicity of spike glycoprotein-based candidate vaccines in the near future. |
topic |
COVID-19 Coronavirus SARS SARS-CoV-2 Neutralization assay Pseudotyped virus |
url |
https://doi.org/10.1186/s12985-020-01472-1 |
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