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An Integrated Research–Clinical BSL-2 Platform for a Live SARS-CoV-2 Neutralization Assay
A reliable and efficient serological test is crucial for monitoring neutralizing antibodies against SARS-CoV-2 and its variants of concern (VOCs). Here, we present an integrated research–clinical platform for a live SARS-CoV-2 neutralization assay, utilizing highly attenuated SARS-CoV-2 (Δ3678_WA1-s...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10536566/ https://www.ncbi.nlm.nih.gov/pubmed/37766263 http://dx.doi.org/10.3390/v15091855 |
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author | Zou, Jing Kurhade, Chaitanya Chang, Hope C. Hu, Yanping Meza, Jose A. Beaver, David Trinh, Ky Omlid, Joseph Elghetany, Bassem Desai, Ragini McCaffrey, Peter Garcia, Juan D. Shi, Pei-Yong Ren, Ping Xie, Xuping |
author_facet | Zou, Jing Kurhade, Chaitanya Chang, Hope C. Hu, Yanping Meza, Jose A. Beaver, David Trinh, Ky Omlid, Joseph Elghetany, Bassem Desai, Ragini McCaffrey, Peter Garcia, Juan D. Shi, Pei-Yong Ren, Ping Xie, Xuping |
author_sort | Zou, Jing |
collection | PubMed |
description | A reliable and efficient serological test is crucial for monitoring neutralizing antibodies against SARS-CoV-2 and its variants of concern (VOCs). Here, we present an integrated research–clinical platform for a live SARS-CoV-2 neutralization assay, utilizing highly attenuated SARS-CoV-2 (Δ3678_WA1-spike). This strain contains mutations in viral transcription regulation sequences and deletion in the open-reading-frames 3, 6, 7, and 8, allowing for safe handling in biosafety level 2 (BSL-2) laboratories. Building on this backbone, we constructed a genetically stable reporter virus (mGFP Δ3678_WA1-spike) by incorporating a modified green fluorescent protein sequence (mGFP). We also constructed mGFP Δ3678_BA.5-spike and mGFP Δ3678_XBB.1.5-spike by substituting the WA1 spike with variants BA.5 and XBB.1.5 spike, respectively. All three viruses exhibit robust fluorescent signals in infected cells and neutralization titers in an optimized fluorescence reduction neutralization assay that highly correlates with a conventional plaque reduction assay. Furthermore, we established that a streamlined robot-aided Bench-to-Clinics COVID-19 Neutralization Test workflow demonstrated remarkably sensitive, specific, reproducible, and accurate characteristics, allowing the assessment of neutralization titers against SARS-CoV-2 variants within 24 h after sample receiving. Overall, our innovative approach provides a valuable avenue for large-scale testing of clinical samples against SARS-CoV-2 and VOCs at BSL-2, supporting pandemic preparedness and response strategies. |
format | Online Article Text |
id | pubmed-10536566 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-105365662023-09-29 An Integrated Research–Clinical BSL-2 Platform for a Live SARS-CoV-2 Neutralization Assay Zou, Jing Kurhade, Chaitanya Chang, Hope C. Hu, Yanping Meza, Jose A. Beaver, David Trinh, Ky Omlid, Joseph Elghetany, Bassem Desai, Ragini McCaffrey, Peter Garcia, Juan D. Shi, Pei-Yong Ren, Ping Xie, Xuping Viruses Article A reliable and efficient serological test is crucial for monitoring neutralizing antibodies against SARS-CoV-2 and its variants of concern (VOCs). Here, we present an integrated research–clinical platform for a live SARS-CoV-2 neutralization assay, utilizing highly attenuated SARS-CoV-2 (Δ3678_WA1-spike). This strain contains mutations in viral transcription regulation sequences and deletion in the open-reading-frames 3, 6, 7, and 8, allowing for safe handling in biosafety level 2 (BSL-2) laboratories. Building on this backbone, we constructed a genetically stable reporter virus (mGFP Δ3678_WA1-spike) by incorporating a modified green fluorescent protein sequence (mGFP). We also constructed mGFP Δ3678_BA.5-spike and mGFP Δ3678_XBB.1.5-spike by substituting the WA1 spike with variants BA.5 and XBB.1.5 spike, respectively. All three viruses exhibit robust fluorescent signals in infected cells and neutralization titers in an optimized fluorescence reduction neutralization assay that highly correlates with a conventional plaque reduction assay. Furthermore, we established that a streamlined robot-aided Bench-to-Clinics COVID-19 Neutralization Test workflow demonstrated remarkably sensitive, specific, reproducible, and accurate characteristics, allowing the assessment of neutralization titers against SARS-CoV-2 variants within 24 h after sample receiving. Overall, our innovative approach provides a valuable avenue for large-scale testing of clinical samples against SARS-CoV-2 and VOCs at BSL-2, supporting pandemic preparedness and response strategies. MDPI 2023-08-31 /pmc/articles/PMC10536566/ /pubmed/37766263 http://dx.doi.org/10.3390/v15091855 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Zou, Jing Kurhade, Chaitanya Chang, Hope C. Hu, Yanping Meza, Jose A. Beaver, David Trinh, Ky Omlid, Joseph Elghetany, Bassem Desai, Ragini McCaffrey, Peter Garcia, Juan D. Shi, Pei-Yong Ren, Ping Xie, Xuping An Integrated Research–Clinical BSL-2 Platform for a Live SARS-CoV-2 Neutralization Assay |
title | An Integrated Research–Clinical BSL-2 Platform for a Live SARS-CoV-2 Neutralization Assay |
title_full | An Integrated Research–Clinical BSL-2 Platform for a Live SARS-CoV-2 Neutralization Assay |
title_fullStr | An Integrated Research–Clinical BSL-2 Platform for a Live SARS-CoV-2 Neutralization Assay |
title_full_unstemmed | An Integrated Research–Clinical BSL-2 Platform for a Live SARS-CoV-2 Neutralization Assay |
title_short | An Integrated Research–Clinical BSL-2 Platform for a Live SARS-CoV-2 Neutralization Assay |
title_sort | integrated research–clinical bsl-2 platform for a live sars-cov-2 neutralization assay |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10536566/ https://www.ncbi.nlm.nih.gov/pubmed/37766263 http://dx.doi.org/10.3390/v15091855 |
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