Cargando…
Evaluation of SARS-CoV-2 neutralizing antibodies using a vesicular stomatitis virus possessing SARS-CoV-2 spike protein
BACKGROUND: SARS-CoV-2 is a novel coronavirus that emerged in 2019 and is now classified in the genus Coronavirus with closely related SARS-CoV. SARS-CoV-2 is highly pathogenic in humans and is classified as a biosafety level (BSL)-3 pathogen, which makes manipulating it relatively difficult due to...
Autores principales: | , , , , , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7801864/ https://www.ncbi.nlm.nih.gov/pubmed/33435994 http://dx.doi.org/10.1186/s12985-021-01490-7 |
_version_ | 1783635661518012416 |
---|---|
author | Tani, Hideki Kimura, Miyuki Tan, Long Yoshida, Yoshihiro Ozawa, Tatsuhiko Kishi, Hiroyuki Fukushi, Shuetsu Saijo, Masayuki Sano, Kaori Suzuki, Tadaki Kawasuji, Hitoshi Ueno, Akitoshi Miyajima, Yuki Fukui, Yasutaka Sakamaki, Ippei Yamamoto, Yoshihiro Morinaga, Yoshitomo |
author_facet | Tani, Hideki Kimura, Miyuki Tan, Long Yoshida, Yoshihiro Ozawa, Tatsuhiko Kishi, Hiroyuki Fukushi, Shuetsu Saijo, Masayuki Sano, Kaori Suzuki, Tadaki Kawasuji, Hitoshi Ueno, Akitoshi Miyajima, Yuki Fukui, Yasutaka Sakamaki, Ippei Yamamoto, Yoshihiro Morinaga, Yoshitomo |
author_sort | Tani, Hideki |
collection | PubMed |
description | BACKGROUND: SARS-CoV-2 is a novel coronavirus that emerged in 2019 and is now classified in the genus Coronavirus with closely related SARS-CoV. SARS-CoV-2 is highly pathogenic in humans and is classified as a biosafety level (BSL)-3 pathogen, which makes manipulating it relatively difficult due to its infectious nature. METHODS: To circumvent the need for BSL-3 laboratories, an alternative assay was developed that avoids live virus and instead uses a recombinant VSV expressing luciferase and possesses the full length or truncated spike proteins of SARS-CoV-2. Furthermore, to measure SARS-CoV-2 neutralizing antibodies under BSL2 conditions, a chemiluminescence reduction neutralization test (CRNT) for SARS-CoV-2 was developed. The neutralization values of the serum samples collected from hospitalized patients with COVID-19 or SARS-CoV-2 PCR-negative donors against the pseudotyped virus infection evaluated by the CRNT were compared with antibody titers determined from an enzyme-linked immunosorbent assay (ELISA) or an immunofluorescence assay (IFA). RESULTS: The CRNT, which used whole blood collected from hospitalized patients with COVID-19, was also examined. As a result, the inhibition of pseudotyped virus infection was specifically observed in both serum and whole blood and was also correlated with the results of the IFA. CONCLUSIONS: In conclusion, the CRNT for COVID-19 is a convenient assay system that can be performed in a BSL-2 laboratory with high specificity and sensitivity for evaluating the occurrence of neutralizing antibodies against SARS-CoV-2. |
format | Online Article Text |
id | pubmed-7801864 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-78018642021-01-12 Evaluation of SARS-CoV-2 neutralizing antibodies using a vesicular stomatitis virus possessing SARS-CoV-2 spike protein Tani, Hideki Kimura, Miyuki Tan, Long Yoshida, Yoshihiro Ozawa, Tatsuhiko Kishi, Hiroyuki Fukushi, Shuetsu Saijo, Masayuki Sano, Kaori Suzuki, Tadaki Kawasuji, Hitoshi Ueno, Akitoshi Miyajima, Yuki Fukui, Yasutaka Sakamaki, Ippei Yamamoto, Yoshihiro Morinaga, Yoshitomo Virol J Research BACKGROUND: SARS-CoV-2 is a novel coronavirus that emerged in 2019 and is now classified in the genus Coronavirus with closely related SARS-CoV. SARS-CoV-2 is highly pathogenic in humans and is classified as a biosafety level (BSL)-3 pathogen, which makes manipulating it relatively difficult due to its infectious nature. METHODS: To circumvent the need for BSL-3 laboratories, an alternative assay was developed that avoids live virus and instead uses a recombinant VSV expressing luciferase and possesses the full length or truncated spike proteins of SARS-CoV-2. Furthermore, to measure SARS-CoV-2 neutralizing antibodies under BSL2 conditions, a chemiluminescence reduction neutralization test (CRNT) for SARS-CoV-2 was developed. The neutralization values of the serum samples collected from hospitalized patients with COVID-19 or SARS-CoV-2 PCR-negative donors against the pseudotyped virus infection evaluated by the CRNT were compared with antibody titers determined from an enzyme-linked immunosorbent assay (ELISA) or an immunofluorescence assay (IFA). RESULTS: The CRNT, which used whole blood collected from hospitalized patients with COVID-19, was also examined. As a result, the inhibition of pseudotyped virus infection was specifically observed in both serum and whole blood and was also correlated with the results of the IFA. CONCLUSIONS: In conclusion, the CRNT for COVID-19 is a convenient assay system that can be performed in a BSL-2 laboratory with high specificity and sensitivity for evaluating the occurrence of neutralizing antibodies against SARS-CoV-2. BioMed Central 2021-01-12 /pmc/articles/PMC7801864/ /pubmed/33435994 http://dx.doi.org/10.1186/s12985-021-01490-7 Text en © The Author(s) 2021 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Tani, Hideki Kimura, Miyuki Tan, Long Yoshida, Yoshihiro Ozawa, Tatsuhiko Kishi, Hiroyuki Fukushi, Shuetsu Saijo, Masayuki Sano, Kaori Suzuki, Tadaki Kawasuji, Hitoshi Ueno, Akitoshi Miyajima, Yuki Fukui, Yasutaka Sakamaki, Ippei Yamamoto, Yoshihiro Morinaga, Yoshitomo Evaluation of SARS-CoV-2 neutralizing antibodies using a vesicular stomatitis virus possessing SARS-CoV-2 spike protein |
title | Evaluation of SARS-CoV-2 neutralizing antibodies using a vesicular stomatitis virus possessing SARS-CoV-2 spike protein |
title_full | Evaluation of SARS-CoV-2 neutralizing antibodies using a vesicular stomatitis virus possessing SARS-CoV-2 spike protein |
title_fullStr | Evaluation of SARS-CoV-2 neutralizing antibodies using a vesicular stomatitis virus possessing SARS-CoV-2 spike protein |
title_full_unstemmed | Evaluation of SARS-CoV-2 neutralizing antibodies using a vesicular stomatitis virus possessing SARS-CoV-2 spike protein |
title_short | Evaluation of SARS-CoV-2 neutralizing antibodies using a vesicular stomatitis virus possessing SARS-CoV-2 spike protein |
title_sort | evaluation of sars-cov-2 neutralizing antibodies using a vesicular stomatitis virus possessing sars-cov-2 spike protein |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7801864/ https://www.ncbi.nlm.nih.gov/pubmed/33435994 http://dx.doi.org/10.1186/s12985-021-01490-7 |
work_keys_str_mv | AT tanihideki evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT kimuramiyuki evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT tanlong evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT yoshidayoshihiro evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT ozawatatsuhiko evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT kishihiroyuki evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT fukushishuetsu evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT saijomasayuki evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT sanokaori evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT suzukitadaki evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT kawasujihitoshi evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT uenoakitoshi evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT miyajimayuki evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT fukuiyasutaka evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT sakamakiippei evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT yamamotoyoshihiro evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein AT morinagayoshitomo evaluationofsarscov2neutralizingantibodiesusingavesicularstomatitisviruspossessingsarscov2spikeprotein |