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The theory and practice of the viral dose in neutralization assay: Insights on SARS-CoV-2 “doublethink” effect
The neutralization assays are considered the gold-standard being capable of evaluating and detecting, functional antibodies. To date, many different protocols exist for micro-neutralization (MN) assay which varies in several steps: cell number and seeding conditions, virus amount used in the infecti...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Author(s). Published by Elsevier B.V.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8364219/ https://www.ncbi.nlm.nih.gov/pubmed/34403775 http://dx.doi.org/10.1016/j.jviromet.2021.114261 |
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author | Manenti, Alessandro Molesti, Eleonora Maggetti, Marta Torelli, Alessandro Lapini, Giulia Montomoli, Emanuele |
author_facet | Manenti, Alessandro Molesti, Eleonora Maggetti, Marta Torelli, Alessandro Lapini, Giulia Montomoli, Emanuele |
author_sort | Manenti, Alessandro |
collection | PubMed |
description | The neutralization assays are considered the gold-standard being capable of evaluating and detecting, functional antibodies. To date, many different protocols exist for micro-neutralization (MN) assay which varies in several steps: cell number and seeding conditions, virus amount used in the infection step, virus-serum-cells incubation time and read out. The aim of the present preliminary study was to carry out SARS-CoV-2 wild type MN assay in order to investigate which optimal tissue culture infective dose 50 (TCID(50)) infective dose in use is the most adequate choice for implementation in terms of reproducibility, standardization possibilities and comparability of results. Therefore, we assessed the MN by using two viral infective doses: the “standard” dose of 100 TCID(50)/well and a reduced dose of 25 TCID(50)/well. The results obtained, yielded by MN on using the lower infective dose (25 TCID50), were higher respect to those obtained with the standard infective dose. This suggests that the lower dose can potentially have a positive impact on the detection and estimation of real amount of neutralizing antibodies present in a given sample, showing higher sensitivity maintaining high specificity. |
format | Online Article Text |
id | pubmed-8364219 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | The Author(s). Published by Elsevier B.V. |
record_format | MEDLINE/PubMed |
spelling | pubmed-83642192021-08-15 The theory and practice of the viral dose in neutralization assay: Insights on SARS-CoV-2 “doublethink” effect Manenti, Alessandro Molesti, Eleonora Maggetti, Marta Torelli, Alessandro Lapini, Giulia Montomoli, Emanuele J Virol Methods Short Communication The neutralization assays are considered the gold-standard being capable of evaluating and detecting, functional antibodies. To date, many different protocols exist for micro-neutralization (MN) assay which varies in several steps: cell number and seeding conditions, virus amount used in the infection step, virus-serum-cells incubation time and read out. The aim of the present preliminary study was to carry out SARS-CoV-2 wild type MN assay in order to investigate which optimal tissue culture infective dose 50 (TCID(50)) infective dose in use is the most adequate choice for implementation in terms of reproducibility, standardization possibilities and comparability of results. Therefore, we assessed the MN by using two viral infective doses: the “standard” dose of 100 TCID(50)/well and a reduced dose of 25 TCID(50)/well. The results obtained, yielded by MN on using the lower infective dose (25 TCID50), were higher respect to those obtained with the standard infective dose. This suggests that the lower dose can potentially have a positive impact on the detection and estimation of real amount of neutralizing antibodies present in a given sample, showing higher sensitivity maintaining high specificity. The Author(s). Published by Elsevier B.V. 2021-11 2021-08-14 /pmc/articles/PMC8364219/ /pubmed/34403775 http://dx.doi.org/10.1016/j.jviromet.2021.114261 Text en © 2021 The Author(s) Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Short Communication Manenti, Alessandro Molesti, Eleonora Maggetti, Marta Torelli, Alessandro Lapini, Giulia Montomoli, Emanuele The theory and practice of the viral dose in neutralization assay: Insights on SARS-CoV-2 “doublethink” effect |
title | The theory and practice of the viral dose in neutralization assay: Insights on SARS-CoV-2 “doublethink” effect |
title_full | The theory and practice of the viral dose in neutralization assay: Insights on SARS-CoV-2 “doublethink” effect |
title_fullStr | The theory and practice of the viral dose in neutralization assay: Insights on SARS-CoV-2 “doublethink” effect |
title_full_unstemmed | The theory and practice of the viral dose in neutralization assay: Insights on SARS-CoV-2 “doublethink” effect |
title_short | The theory and practice of the viral dose in neutralization assay: Insights on SARS-CoV-2 “doublethink” effect |
title_sort | theory and practice of the viral dose in neutralization assay: insights on sars-cov-2 “doublethink” effect |
topic | Short Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8364219/ https://www.ncbi.nlm.nih.gov/pubmed/34403775 http://dx.doi.org/10.1016/j.jviromet.2021.114261 |
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