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Neutralization assay with SARS-CoV-1 and SARS-CoV-2 spike pseudotyped murine leukemia virions.
Virol J. 2021 01 04; 18(1):1.VJ

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.

Authors+Show Affiliations

Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA.Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA.Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA.RetroVirox, Inc., San Diego, CA, USA.RetroVirox, Inc., San Diego, CA, USA.Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA. Associated Regional and University Pathologists (ARUP) Laboratories, Salt Lake City, UT, USA.Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA. Associated Regional and University Pathologists (ARUP) Laboratories, Salt Lake City, UT, USA.Associated Regional and University Pathologists (ARUP) Laboratories, Salt Lake City, UT, USA.Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA.AIDS Immunopathology Unit, National Center of Microbiology (CNM), Instituto de Salud Carlos III, Madrid, Spain.Department of Human Genetics, University of Utah School of Medicine, Salt Lake City, UT, USA.Department of Medicine, University of Utah School of Medicine, Salt Lake City, UT, USA.Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT, USA. vicente.planelles@path.utah.edu.

Pub Type(s)

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

Language

eng

PubMed ID

33397387

Citation

Zheng, Yue, et al. "Neutralization Assay With SARS-CoV-1 and SARS-CoV-2 Spike Pseudotyped Murine Leukemia Virions." Virology Journal, vol. 18, no. 1, 2021, p. 1.
Zheng Y, Larragoite ET, Williams ESCP, et al. Neutralization assay with SARS-CoV-1 and SARS-CoV-2 spike pseudotyped murine leukemia virions. Virol J. 2021;18(1):1.
Zheng, Y., Larragoite, E. T., Williams, E. S. C. P., Lama, J., Cisneros, I., Delgado, J. C., Slev, P., Rychert, J., Innis, E. A., Coiras, M., Rondina, M. T., Spivak, A. M., & Planelles, V. (2021). Neutralization assay with SARS-CoV-1 and SARS-CoV-2 spike pseudotyped murine leukemia virions. Virology Journal, 18(1), 1. https://doi.org/10.1186/s12985-020-01472-1
Zheng Y, et al. Neutralization Assay With SARS-CoV-1 and SARS-CoV-2 Spike Pseudotyped Murine Leukemia Virions. Virol J. 2021 01 4;18(1):1. PubMed PMID: 33397387.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Neutralization assay with SARS-CoV-1 and SARS-CoV-2 spike pseudotyped murine leukemia virions. AU - Zheng,Yue, AU - Larragoite,Erin T, AU - Williams,Elizabeth S C P, AU - Lama,Juan, AU - Cisneros,Isabel, AU - Delgado,Julio C, AU - Slev,Patricia, AU - Rychert,Jenna, AU - Innis,Emily A, AU - Coiras,Mayte, AU - Rondina,Matthew T, AU - Spivak,Adam M, AU - Planelles,Vicente, Y1 - 2021/01/04/ PY - 2020/09/15/received PY - 2020/12/16/accepted PY - 2021/1/5/entrez PY - 2021/1/6/pubmed PY - 2021/1/15/medline KW - Antibody KW - COVID-19 KW - Coronavirus KW - Murine leukemia virus KW - Neutralization assay KW - Pseudotyped virus KW - SARS KW - SARS-CoV-2 KW - Spike SP - 1 EP - 1 JF - Virology journal JO - Virol J VL - 18 IS - 1 N2 - 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. SN - 1743-422X UR - https://www.unboundmedicine.com/medline/citation/33397387/Neutralization_assay_with_SARS_CoV_1_and_SARS_CoV_2_spike_pseudotyped_murine_leukemia_virions_ L2 - https://virologyj.biomedcentral.com/articles/10.1186/s12985-020-01472-1 DB - PRIME DP - Unbound Medicine ER -