tetano
Editor, Senior Moderator
J Med Virol. 2020 May 8. doi: 10.1002/jmv.25986. [Epub ahead of print]
Evaluation of SARS-CoV-2 neutralizing antibodies using of a CPE-based Colorimetric live virus micro-neutralization assay in human serum samples.
Manenti A[SUP]1,[/SUP][SUP]2[/SUP], Maggetti M[SUP]2[/SUP], Casa E[SUP]1,[/SUP][SUP]2[/SUP], Martinuzzi D[SUP]1[/SUP], Torelli A[SUP]1[/SUP], Trombetta CM[SUP]3[/SUP], Marchi S[SUP]3[/SUP], Montomoli E[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP].
Author information
Abstract
The micro-neutralization assay is a fundamental test in virology, immunology, vaccine assessment and epidemiology studies. Since the SARS-CoV-2 outbreak at the end of December 2019 in China, it has become extremely important to have well-established and validated diagnostic and serological assays for this new emerging virus. Here, we present a micro-neutralization assay with the use of SARS-CoV-2 wild type virus with two different methods of read-out. We evaluated the performance of this assay using human serum samples taken from an italian seroepidemiological study being performed at the University of Siena, along with the human monoclonal antibody CR3022 and some iper-immune animal serum samples against Influenza and Adenovirus strains. The same panel of human samples have been previously tested in ELISA as a pre-screening. Positive, borderline and negative ELISA samples were evaluated in neutralization assay using two different method of read-out: subjective (by means of an inverted optical microscope) and objective (by means of a spectrophotometer). Our findings suggest that at least 50% of positive ELISA samples are positive in neutralization as well, and that method is able to quantify different antibody concentrations in a specific manner. Taken together, our results confirm that the colorimetric cytopathic effect-based microneutralization assay could be used as a valid clinical test method for epidemiological and vaccines studies. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
KEYWORDS:
Neutralization; SARS Coronavirus; epidemiology; humoral immunity; pandemic
PMID:32383254DOI:10.1002/jmv.25986
Evaluation of SARS-CoV-2 neutralizing antibodies using of a CPE-based Colorimetric live virus micro-neutralization assay in human serum samples.
Manenti A[SUP]1,[/SUP][SUP]2[/SUP], Maggetti M[SUP]2[/SUP], Casa E[SUP]1,[/SUP][SUP]2[/SUP], Martinuzzi D[SUP]1[/SUP], Torelli A[SUP]1[/SUP], Trombetta CM[SUP]3[/SUP], Marchi S[SUP]3[/SUP], Montomoli E[SUP]1,[/SUP][SUP]2,[/SUP][SUP]3[/SUP].
Author information
Abstract
The micro-neutralization assay is a fundamental test in virology, immunology, vaccine assessment and epidemiology studies. Since the SARS-CoV-2 outbreak at the end of December 2019 in China, it has become extremely important to have well-established and validated diagnostic and serological assays for this new emerging virus. Here, we present a micro-neutralization assay with the use of SARS-CoV-2 wild type virus with two different methods of read-out. We evaluated the performance of this assay using human serum samples taken from an italian seroepidemiological study being performed at the University of Siena, along with the human monoclonal antibody CR3022 and some iper-immune animal serum samples against Influenza and Adenovirus strains. The same panel of human samples have been previously tested in ELISA as a pre-screening. Positive, borderline and negative ELISA samples were evaluated in neutralization assay using two different method of read-out: subjective (by means of an inverted optical microscope) and objective (by means of a spectrophotometer). Our findings suggest that at least 50% of positive ELISA samples are positive in neutralization as well, and that method is able to quantify different antibody concentrations in a specific manner. Taken together, our results confirm that the colorimetric cytopathic effect-based microneutralization assay could be used as a valid clinical test method for epidemiological and vaccines studies. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
KEYWORDS:
Neutralization; SARS Coronavirus; epidemiology; humoral immunity; pandemic
PMID:32383254DOI:10.1002/jmv.25986