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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...

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Autores principales: Manenti, Alessandro, Molesti, Eleonora, Maggetti, Marta, Torelli, Alessandro, Lapini, Giulia, Montomoli, Emanuele
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Author(s). Published by Elsevier B.V. 2021
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.
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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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